Protein detection

Last updated

Protein detection is used for clinical diagnosis, treatment and biological research. [1] Protein detection evaluates the concentration and amount of different proteins in a particular specimen. [2] There are different methods and techniques to detect protein in different organisms. Protein detection has demonstrated important implications for clinical diagnosis, treatment and biological research. [3] Protein detection technique has been utilized to discover protein in different category food, such as soybean (bean), walnut (nut), and beef (meat). [4] Protein detection method for different type food vary on the basis of property of food for bean, nut and meat. Protein detection has different application in different field.

Contents

Protein Detection in Soybeans, Walnuts, Beef

Detection of Functional Modes in Protein

Purpose for protein detection in food

Soybean Plant Soybeans.jpg
Soybean Plant

Allergies from food have been noted to become common disease nowadays. The food allergies in the clinical demonstration present different signs, for example mild symptoms from itching in the mouth and swelling of the lips to critical anaphylactic response result in fatal consequences. [5] According to statistic, about 2% adults and 8% children are experiencing hypersensitivity from industrialized countries. In order to reduce potential threatening reactions for life, avoiding the consumption from these allergenic foods strictly is  the valid therapy. Therefore, sufficient description in term of potentially allergenic ingredients existing in food products is crucial and indispensable which can be monitored through protein detection. [6] [7]

Rationale for protein detection in soybeans

The soybean has been consumed in processed foods all over the word because of its high nutrient and easy processing characteristic such as soybean milk, tofu, meat alternatives, and brewed soybean products. [5] microorganisms is used in brewage process for brewed soybean products like miso, soy sauce, natto and tempeh. Allergenicity stays in brewed soybean products. In Asian countries, these brewed soybean products are popular and traditional. The amount of patients from soybean allergy and the nearly infinite uses for soybean have gone up in the past a couple of years. [8]

Previous method for protein detection in soybeans

During the last 30 years, broad methods and techniques were experimented to discover soybean protein. These methods and techniques can be conveyed to lab environment easily. [9] The original and traditional methods were designed and tested in molecular biology spectrum. Enzyme‐Linked Immunosorbent Assay technique containing high susceptibility and specificity is reliable method to investigate soybean proteins through applying a protein which can identify a foreign molecule. This has been evaluated as a vacuolar protein including a molecular block of 34 kDa. The ELISA illustrated sufficient repeatability and reproducibility in lab assessment. But it can not test protein in soybean existing in brewed soybean products. [10] There are different studies to conduct experiments to assess soybean protein through ELISA. However, reproducibility, cross-reactivity and low repeatability make measurement difficult to be reliable in processed foods. These methods can not discover soybean protein staying in brewed soybean products. [11]

Current method for protein detection in soybeans

Compared with previous method, a heating process is involved in current abstraction technique to investigate soybean protein existing in brewed products. Since the heating process can deactivate the microbial proteolytic enzymes, the current abstraction technique can be used to disclose soybean protein in brewed soybean products. [12] The heating abstraction technique can be demonstrated as the following. To produce the good dispersibility for the specimen in the extraction buffer to carry out the heating process, 19mL of abstraction buffer is mixed with five glass beads in five millimeter diameter and 1 g of food homogenate. At 5, 15 and 60 min variable time, the mixture is abstracted under 25, 40, 60, 80 and 100 ° variable temperature through the heating in a water bath followed by every 5 minutes vortexing. Food abstractions generated through the previous and the current technique are centrifuged for 20 minutes at three thousand gram, then the supernatant is filtered off by a filter paper. The filtrate is gathered and applied for analysis immediately acting as the food specimen abstract. [13] The calibration standard solutions needs to be prepared to disclose soybean proteins by using ELISA.  A three hundred milligram soybean powder specimen is mixed with a twenty milliliter compound including 0.5 M NaCl, 0.5% SDS, 20 mM Tris-HCl (pH 7.5), and 2% 2-ME. The compound is then shaken at room temperature for 16 hours for abstraction. The abstract is centrifuged for 30 minutes at twenty thousand gram, then the supernatant is selected by a 0.8-μm microfilter paper. The protein substance from the initial abstract is inspected with a 2-D Quant Kit. The initial abstract is diluted to 50 ng/mL combined with 0.1% SDS, 0.1% 2-ME, 0.1 M PBS (pH 7.4), 0.1% BSA, and 0.1% Tween 20, and it is deposited for ELISA at 4 °C playing as the calibration standard solution. [8]

Conclusion for current protein detection method in soybeans.

The detection limit for the ELISA is 1 μg/g and it can not assess soybean proteins existing in brewed soybean products due to degradation of the proteins in soybean through microbial proteolytic enzymes staying in the brewed products. The microbial proteolytic enzymes possibly restrain the detection of soybean protein storing in the brewed soybean products. The current abstraction technique can control protein degradation through the microbial proteolytic enzymes. The microbial proteolytic enzymes can be inhibited by heating, pH, and protease inhibitors in general. [14] The variable heating temperatures and abstraction times are examined to decide the ideal heating temperature and time to control microbial proteolytic enzymes. The heating conditions showed to optimize the control of microbial proteolytic enzymes is 80 °C for 15 minutes. So the heating temperature for the abstraction is set to 80 °C and the time is set to 15 minutes for the current abstraction technique. [15]

The current abstraction technique can restrain the degradation of soybean proteins through microbial proteolytic enzymes and can detect soybean protein in most brewed soybean products. The current abstraction technique combined with the heating is a useful and sensitive tool to discover soybean protein stored in processed foods and brewed soybean products. Without impacting microbial proteolytic enzymes, this method is appropriate to quantify soybean protein in processed foods. The proposed extraction and ELISA technique can be applied to control labeling systems for soybean ingredient through a trusty manner. [8]

Black Walnut Black Walnut nut and leave detail.JPG
Black Walnut
English Walnut Juglans regia 2009 G2.jpg
English Walnut

Rationale for protein detection in walnuts

English walnuts (Juglans regia) and black walnuts (Juglans nigra) are two main types of walnuts in the market across the world. Walnuts are utilized as a valuable ingredient due to favorable health attributes, sensory properties and consumer sensation. [16] [17] Shelled walnuts are broadly applied as ingredients in different foods such as salad, ice creams, bread and meat alternative. Walnut oil is introduced as a good source of mono- and polyunsaturated fatty acids and tocopherols. [18] And it is adopted as a food ingredient in salad dressings particularly. Walnut hull extract is considered as a dietary supplement and a seasoning in the food industry. In addition, ground walnut shells can be used in industrial field as extenders, carriers, fillers and abrasives for example jet cleaners. Tree nuts are regarded as one of the most common allergenic foods around the world. [19]   Allergic reactions from tree nuts can be fierce and life threatening. [20] [21] Individuals with walnut allergies can have result in fatal and near-fatal reactions from the unintended ingestion of walnuts, other tree nuts or possibly contamination of food with the walnuts ingredient. [22] [23] [24] [25] [26] [27] [28] [29] [30] [31] To prevent walnut allergic reactions, the only effective way is to avoid walnuts in the diet. [32] The appropriate labeling of processed foods with walnuts ingredient is critical to protect walnut-allergic consumers. There are a couple of circumstances cause undeclared walnut residues such as sharing equipment between walnut-containing and other formulations and undeclared walnuts in ingredients. [33] The enzyme-linked immunosorbent assay (ELISA) can be used as the technique to detect walnuts residues with great sensitivity and specificity since walnuts allergic Individuals can have allergic reactions with low (milligram) amounts of walnuts. [34] Several different techniques can be applied to discover walnut residues as well such as polymerase chain reaction (PCR) method and ELISA method on the basis of polyclonal antisera raised against a particular 2S albumin walnut protein. [35] [36] [37]

Current method for protein detection in walnuts

The sandwich-type walnut ELISA is the current method used to detect protein in walnuts. The sandwich-type walnut ELISA can be applied as a critical analytical technique by food manufacturers and regulatory agencies for hygiene validation and the assessment of allergen control strategies.

Immunogen preparation

A mixture of several brands of English walnuts are used to produce the immunogen. The mixed walnuts need to be washed by deionized distilled water 6 times and air-dried. Portion of the walnuts are dry-roasted for 10 minutes at 270 ◦F. The roasted or raw walnuts are cleaved, frozen, and ground to a refined particle size through the blender. The ground roasted and ground raw walnuts are defatted and filtered. Then, the powdered raw or roasted walnuts are air-dried thoroughly. Both the defatted, powdered raw, and roasted walnuts can be utilized as immunogens. Protein concentrations of the defatted powdered immunogens are set through the Kjeldahl method with 46.4% raw defatted walnut and 34.9% roasted defatted walnut. [37]

Polyclonal antibody production and titer determination

Polyclonal antibodies are generated in 1 sheep, 1 goat, and 3 New Zealand white rabbits with each immunogen. The initial subcutaneous injections are given to the 10 animals including 3 rabbits, 1 sheep, and 1 goat on multiple sites with the defatted powdered immunogen and Freunds Complete Adjuvant. Titer values of collected antisera are evaluated by a noncompetitive ELISA method with walnut protein from abstracts of the proper raw or roasted immunogen. [37]

Cross-reactivity study and ELISA method

A variety of tree nuts, seeds, legumes, fruits and food ingredients are assessed for cross-reactivity in the walnut ELISA assay. The modified sandwich ELISA can be used to detect walnuts residues with sheep antiroasted walnut and rabbit antiroasted walnut antisera used as the capture and detector antibodies respectively. [37]

Conclusion for current protein detection method in walnuts

Walnut residues can be disclosed at 1 ppm quantitation limit in a diversity of food such as ice cream, muffins, cookies and chocolate. The walnut ELISA can be conducted to detect possible walnut residues allergy in other foods from sharing equipment and to evaluate the sanitation procedures targeted on removal of walnut residues from shared equipment in the food industry. [37]

Cattle Mafate cattle dsc00749.jpg
Cattle

Rationale for protein detection in beef

It has been reported that animal feedingstuffs containing processed animal protein (PAP) contaminated with prions have caused BSE infection of the cattle. Processed animal proteins (PAP) has been prohibited to apply as feed material for all farmed animals except fish meal currently. In addition, infections from consumption of undercooked raw beef has been declaimed to be an important pathogen for Enterohemorrhagic Escherichia coli O157:H7. [38]

Method for protein detection in beef

For processed animal protein, the specific polymerase chain reaction (PCR) based procedure parallelled with microscopic method is utilized to detect processed animal protein (PAP) in feedingstuffs. The limit detection for PCR has been evaluated on 0.05% for beef, 0.1% for pork and 0.2% for poultry meat and bone meal. Microscopic method can disclose 66.13% doubtful samples of feedingstuffs. Combined the results from the use of the microscopic and PCR methods, it has been stated that the molecular biology methods can be executed as a supplementary method for PAP detection. [39]

For undercooked raw beef, in order to make sure a safe beef supply, sensitive and quick detection techniques for E. coli O157:H7 are important in the meat industry. Three different techniques can be used in raw ground beef: the VIDAS ultraperformance E. coli test (ECPT UP), a noncommercial real-time (RT) PCR method and the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) reference method to detect E. coli O157:H7. 25 g of individual raw beef samples and 375 g of raw beef composites can be examined for optimal enrichment times and the efficacy of testing. 6 hours of enrichment is sufficient for both the VIDAS ECPT UP and RT-PCR methods for 25 g samples of each type of raw ground beef, but 24 hours of enrichment is acquired for 375 g samples, Both the VIDAS ECPT UP and RT-PCR methods can generate similar results with those gained from the USDA-FSIS reference method after 18 to 24 hours of enrichment. Low levels of E. coli O157:H7 in 25 g of various types of raw ground beef can be disclosed through these methods, E. coli O157:H7 in composite raw ground beef up to 375 g can be detected as well. [38]

Implication from protein detection

Protein detection in cells from the human rectal mucous membrane can imply colorectal disease such as colon tumours, inflammatory bowel disease. [40] Protein detection based on antibody microarrays can implicate life signature for example organics and biochemical compounds in the solar system in astrobiology field. [41] Protein detection can monitor soybean protein labeling system in processed foods to protect consumers in a reliable way. [8] The labeling for soybean protein declaimed by protein detection has indicated to be the most important solution. [8] Detailed labeling description for the soybean ingredients in refined foods is required to protect the consumer. [8]

Related Research Articles

<span class="mw-page-title-main">Beef</span> Meat from cattle

Beef is the culinary name for meat from cattle. Beef can be prepared in various ways; cuts are often used for steak, which can be cooked to varying degrees of doneness, while trimmings are often ground or minced, as found in most hamburgers. Beef contains protein, iron, and vitamin B12. Along with other kinds of red meat, high consumption is associated with an increased risk of colorectal cancer and coronary heart disease, especially when processed. Beef has a high environmental impact, being a primary driver of deforestation with the highest greenhouse gas emissions of any agricultural product.

<span class="mw-page-title-main">Protease</span> Enzyme that cleaves other proteins into smaller peptides

A protease is an enzyme that catalyzes proteolysis, breaking down proteins into smaller polypeptides or single amino acids, and spurring the formation of new protein products. They do this by cleaving the peptide bonds within proteins by hydrolysis, a reaction where water breaks bonds. Proteases are involved in numerous biological pathways, including digestion of ingested proteins, protein catabolism, and cell signaling.

<span class="mw-page-title-main">Soybean</span> Legume grown for its edible bean

The soybean, soy bean, or soya bean is a species of legume native to East Asia, widely grown for its edible bean, which has numerous uses.

<span class="mw-page-title-main">Casein</span> Family of proteins found in milk

Casein is a family of related phosphoproteins that are commonly found in mammalian milk, comprising about 80% of the proteins in cow's milk and between 20% and 60% of the proteins in human milk. Sheep and cow milk have a higher casein content than other types of milk with human milk having a particularly low casein content.

<span class="mw-page-title-main">ELISA</span> Method to detect an antigen using an antibody and enzyme

The enzyme-linked immunosorbent assay (ELISA) is a commonly used analytical biochemistry assay, first described by Eva Engvall and Peter Perlmann in 1971. The assay is a solid-phase type of enzyme immunoassay (EIA) to detect the presence of a ligand in a liquid sample using antibodies directed against the protein to be measured. ELISA has been used as a diagnostic tool in medicine, plant pathology, and biotechnology, as well as a quality control check in various industries.

<span class="mw-page-title-main">Plant pathology</span> Scientific study of plant diseases

Plant pathology or phytopathology is the scientific study of plant diseases caused by pathogens and environmental conditions. Plant pathology involves the study of pathogen identification, disease etiology, disease cycles, economic impact, plant disease epidemiology, plant disease resistance, how plant diseases affect humans and animals, pathosystem genetics, and management of plant diseases.

<span class="mw-page-title-main">Immunostaining</span> Biochemical technique

In biochemistry, immunostaining is any use of an antibody-based method to detect a specific protein in a sample. The term "immunostaining" was originally used to refer to the immunohistochemical staining of tissue sections, as first described by Albert Coons in 1941. However, immunostaining now encompasses a broad range of techniques used in histology, cell biology, and molecular biology that use antibody-based staining methods.

<span class="mw-page-title-main">Immunoassay</span> Biochemical test for a protein or other molecule using an antibody

An immunoassay (IA) is a biochemical test that measures the presence or concentration of a macromolecule or a small molecule in a solution through the use of an antibody (usually) or an antigen (sometimes). The molecule detected by the immunoassay is often referred to as an "analyte" and is in many cases a protein, although it may be other kinds of molecules, of different sizes and types, as long as the proper antibodies that have the required properties for the assay are developed. Analytes in biological liquids such as serum or urine are frequently measured using immunoassays for medical and research purposes.

<span class="mw-page-title-main">Zymography</span> Electrophoretic hydrolytic enzyme detection technique

Zymography is an electrophoretic technique for the detection of hydrolytic enzymes, based on the substrate repertoire of the enzyme. Three types of zymography are used; in gel zymography, in situ zymography and in vivo zymography. For instance, gelatin embedded in a polyacrylamide gel will be digested by active gelatinases run through the gel. After Coomassie staining, areas of degradation are visible as clear bands against a darkly stained background.

<span class="mw-page-title-main">Food microbiology</span> Study of the microorganisms that inhibit, create, or contaminate food

Food microbiology is the study of the microorganisms that inhabit, create, or contaminate food. This includes the study of microorganisms causing food spoilage; pathogens that may cause disease ; microbes used to produce fermented foods such as cheese, yogurt, bread, beer, and wine; and microbes with other useful roles, such as producing probiotics.

<span class="mw-page-title-main">Soy protein</span> A protein that is isolated from soybean

Soy protein is a protein that is isolated from soybean. It is made from soybean meal that has been dehulled and defatted. Dehulled and defatted soybeans are processed into three kinds of high protein commercial products: soy flour, concentrates, and isolates. Soy protein isolate has been used since 1959 in foods for its functional properties.

<span class="mw-page-title-main">United States raw milk debate</span>

The United States raw milk debate concerns issues of food safety and claimed health benefits of raw milk, and whether authorities responsible for regulating food safety should prohibit sale of raw milk for consumption.

Biomolecular engineering is the application of engineering principles and practices to the purposeful manipulation of molecules of biological origin. Biomolecular engineers integrate knowledge of biological processes with the core knowledge of chemical engineering in order to focus on molecular level solutions to issues and problems in the life sciences related to the environment, agriculture, energy, industry, food production, biotechnology and medicine.

<span class="mw-page-title-main">Soybean meal</span> Ground soybeans used for food

Soybean meal is used in food and animal feeds, principally as a protein supplement, but also as a source of metabolizable energy. Typically 1 bushel of soybeans yields 48 lbs. (21.8 kg) of soybean meal. Soybean meal is produced as a co-product of soybean oil extraction. Some, but not all, soybean meal contains ground soybean hulls. Soybean meal is heat-treated during production, to denature the trypsin inhibitors of soybeans, which would otherwise interfere with protein digestion.

Pathatrix is a high volume recirculating immuno magnetic-capture system developed by Thermo Fisher Scientific for the detection of pathogens in food and environmental samples.

Microbial toxins are toxins produced by micro-organisms, including bacteria, fungi, protozoa, dinoflagellates, and viruses. Many microbial toxins promote infection and disease by directly damaging host tissues and by disabling the immune system. Endotoxins most commonly refer to the lipopolysaccharide (LPS) or lipooligosaccharide (LOS) that are in the outer plasma membrane of Gram-negative bacteria. The botulinum toxin, which is primarily produced by Clostridium botulinum and less frequently by other Clostridium species, is the most toxic substance known in the world. However, microbial toxins also have important uses in medical science and research. Currently, new methods of detecting bacterial toxins are being developed to better isolate and understand these toxins. Potential applications of toxin research include combating microbial virulence, the development of novel anticancer drugs and other medicines, and the use of toxins as tools in neurobiology and cellular biology.

Raw meat generally refers to any type of uncooked muscle tissue of an animal used for food. In the meat production industry, the term ‘meat’ refers specifically to mammalian flesh, while the words ‘poultry’ and ‘seafood’ are used to differentiate between the tissue of birds and aquatic creatures.

Microbial food cultures are live bacteria, yeasts or moulds used in food production. Microbial food cultures carry out the fermentation process in foodstuffs. Used by humans since the Neolithic period fermentation helps to preserve perishable foods and to improve their nutritional and organoleptic qualities. As of 1995, fermented food represented between one quarter and one third of food consumed in Central Europe. More than 260 different species of microbial food culture are identified and described for their beneficial use in fermented food products globally, showing the importance of their use.

<span class="mw-page-title-main">Ara h1</span>

Ara h 1 is a seed storage protein from Arachis hypogaea (peanuts). It is a heat stable 7S vicilin-like globulin with a stable trimeric form that comprises 12-16% of the total protein in peanut extracts. Ara h 1 is known because sensitization to it was found in 95% of peanut-allergic patients from North America. In spite of this high percentage, peanut-allergic patients of European populations have fewer sensitizations to Ara h 1.

Diagnostic microbiology is the study of microbial identification. Since the discovery of the germ theory of disease, scientists have been finding ways to harvest specific organisms. Using methods such as differential media or genome sequencing, physicians and scientists can observe novel functions in organisms for more effective and accurate diagnosis of organisms. Methods used in diagnostic microbiology are often used to take advantage of a particular difference in organisms and attain information about what species it can be identified as, which is often through a reference of previous studies. New studies provide information that others can reference so that scientists can attain a basic understanding of the organism they are examining.

References

  1. Engineering the bioelectronic interface : applications to analyte biosensing and protein detection. Davis, Jason J., Royal Society of Chemistry (Great Britain). Cambridge, UK: RSC Pub. 2009. ISBN   9781615836932. OCLC   701819884.{{cite book}}: CS1 maint: others (link)
  2. "Protein Detection", Electrophoresis in Practice, Wiley-VCH Verlag GmbH & Co. KGaA, 2016-02-26, pp. 131–164, doi:10.1002/9783527695188.ch6, ISBN   9783527695188
  3. Zhang, Hongquan; Li, Feng; Dever, Brittany; Wang, Chuan; Li, Xing-Fang; Le, X. Chris (2013-10-04). "Assembling DNA through Affinity Binding to Achieve Ultrasensitive Protein Detection". Angewandte Chemie International Edition. 52 (41): 10698–10705. doi:10.1002/anie.201210022. PMID   24038633.
  4. Liu, Bin; Teng, Da; Wang, Xiumin; Wang, Jianhua (2013-01-30). "Detection of the Soybean Allergenic Protein Gly m Bd 28K by an Indirect Enzyme-Linked Immunosorbent Assay". Journal of Agricultural and Food Chemistry. 61 (4): 822–828. doi:10.1021/jf303076w. ISSN   0021-8561. PMID   23317377.
  5. 1 2 Wang, Xiaoyu; Jiang, Xiaofeng; Zhu, Shuxian; Liu, Lu; Xia, Junhan; Li, Lidong (2017). "Preparation of optical functional composite films and their application in protein detection". Colloids and Surfaces A: Physicochemical and Engineering Aspects. 535: 69–74. doi:10.1016/j.colsurfa.2017.09.026.
  6. Cheng, Lin; Zhang, Jie; Lin, Yan; Wang, Qiong; Zhang, XiuXiu; Ding, YanHua; Cui, Hanfeng; Fan, Hao (2015). "An electrochemical molecular recognition-based aptasensor for multiple protein detection". Analytical Biochemistry. 491: 31–36. doi:10.1016/j.ab.2015.08.023. PMID   26344894.
  7. Shimojo, Naoshi; Nakamura, Masashi; Sato, Nayu; Sano, Akiyo; Kobayashi, Tsukane; Yagami, Akiko; Kojima, Atsushi; Matsunaga, Kayoko (2016). "Utility of Immunoproteomics in Soybean Allergy". Journal of Allergy and Clinical Immunology. 137 (2): AB139. doi: 10.1016/j.jaci.2015.12.587 .
  8. 1 2 3 4 5 6 Morishita, Naoki; Matsumoto, Takashi; Morimatsu, Fumiki; Toyoda, Masatake (2014). "Detection of Soybean Proteins in Fermented Soybean Products by Using Heating Extraction: Detection of soybean in fermented food…". Journal of Food Science. 79 (5): T1049–T1054. doi:10.1111/1750-3841.12461. PMID   24811351.
  9. Protein blotting and detection : methods and protocols. Kurien, Biji T., Scofield, R. Hal. New York: Humana Press. 2009. ISBN   9781597455428. OCLC   371501294.{{cite book}}: CS1 maint: others (link)
  10. Morales-Narváez, Eden; Guix, Maria; Medina-Sánchez, Mariana; Mayorga-Martinez, Carmen C.; Merkoçi, Arben (2014). "Micromotor Enhanced Microarray Technology for Protein Detection". Small. 10 (13): 2542–2548. doi:10.1002/smll.201303068. hdl: 10261/126975 . PMID   24634101.
  11. Detection of blotted proteins : methods and protocols. Kurien, Biji T.,, Scofield, R. Hal. New York, NY. ISBN   9781493927180. OCLC   913123725.{{cite book}}: CS1 maint: others (link)
  12. Lin, Chenxiang; Katilius, Evaldas; Liu, Yan; Zhang, Junping; Yan, Hao (2006-08-11). "Self-Assembled Signaling Aptamer DNA Arrays for Protein Detection". Angewandte Chemie International Edition. 45 (32): 5296–5301. doi:10.1002/anie.200600438. ISSN   1433-7851. PMID   16847867.
  13. Park, Do Hyun; Lee, Jae-Seung (2015). "Functionalized nanoparticle probes for protein detection". Electronic Materials Letters. 11 (3): 336–345. Bibcode:2015EML....11..336P. doi:10.1007/s13391-014-4383-0. ISSN   1738-8090. S2CID   52949902.
  14. Nong, Rachel Yuan; Gu, Jijuan; Darmanis, Spyros; Kamali-Moghaddam, Masood; Landegren, Ulf (2012). "DNA-assisted protein detection technologies". Expert Review of Proteomics. 9 (1): 21–32. doi:10.1586/epr.11.78. ISSN   1478-9450. PMID   22292821. S2CID   207212401.
  15. Detection of highly dangerous pathogens : microarray methods for the detection of BSL 3 and BSL 4 agents. Kostic, Tanja., Butaye, Patrick., Schrenzel, Jacques. Weinheim: Wiley-VCH. 2009. ISBN   9783527626687. OCLC   463436671.{{cite book}}: CS1 maint: others (link)
  16. Ros, E; Nunez, I; Perez-Heras, A (2004). "A walnut diet improves endothelial function in hypercholesterolemic subjects. A randomized crossover trial". ACC Current Journal Review. 13 (6): 19–20. doi:10.1016/j.accreview.2004.06.062.
  17. Almoosawi, Suzana Fyfe, Lorna Ho, Clement Al-Dujaili, Emad A S (2009-10-13). "The effect of polyphenol-rich dark chocolate on fasting capillary whole blood glucose, total cholesterol, blood pressure and glucocorticoids in healthy overweight and obese subjects". The British Journal of Nutrition. 103 (6). Cambridge University Press: 842–850. doi: 10.1017/S0007114509992431 . OCLC   706594347. PMID   19825207. S2CID   4113907.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  18. Savage, G. P.; Dutta, P. C.; McNeil, D. L. (1999). "Fatty acid and tocopherol contents and oxidative stability of walnut oils". Journal of the American Oil Chemists' Society. 76 (9): 1059–1063. doi:10.1007/s11746-999-0204-2. S2CID   82106409.
  19. Preparation and use of food-based dietary guidelines : report of a Joint FAO/WHO Consultation. Joint FAO/WHO Consultation on the Preparation and Use of Food-Based Dietary Guideline., World Health Organization. Geneva: World Health Organization. 1998. ISBN   9241208805. OCLC   40216171.{{cite book}}: CS1 maint: others (link)
  20. Silver, M.; Comline, R. S. (1975). "Transfer of gases and metabolites in the equine placenta: a comparison with other species". Journal of Reproduction and Fertility. Supplement (23): 589–594. ISSN   0449-3087. PMID   1529.
  21. Teuber, Suzanne S.; Comstock, Sarah S.; Sathe, Shridhar K.; Roux, Kenneth H. (2003). "Tree nut allergy". Current Allergy and Asthma Reports. 3 (1): 54–61. doi:10.1007/s11882-003-0013-x. ISSN   1529-7322. PMID   12542995. S2CID   42075188.
  22. Asthma '84: Pharmacologic Update (1984 : Rancho Mirage, Calif.) (1985). Asthma '84: Pharmacologic Update : October 31 to November 3, 1984, Rancho Mirage, California. Mosby. OCLC   12425311.{{cite book}}: CS1 maint: numeric names: authors list (link)
  23. Bock, S.Allan; Muñoz-Furlong, Anne; Sampson, Hugh A. (2001). "Fatalities due to anaphylactic reactions to foods". Journal of Allergy and Clinical Immunology. 107 (1): 191–193. doi: 10.1067/mai.2001.112031 . PMID   11150011.
  24. Lawrence Berkeley National Laboratory. United States. Department of Energy. Office of Scientific and Technical Information. (2008). Decreased expression of RNA interference machinery, Dicer and Drosha, is associated with poor outcome in ovarian cancer patients. Lawrence Berkeley National Laboratory. OCLC   727220807.
  25. Sampson, Hugh A.; Mendelson, Louis; Rosen, James P. (1992-08-06). "Fatal and Near-Fatal Anaphylactic Reactions to Food in Children and Adolescents". New England Journal of Medicine. 327 (6): 380–384. doi: 10.1056/NEJM199208063270603 . ISSN   0028-4793. PMID   1294076.
  26. Baker, D.G. (1997). Relationship between posttraumatic stress disorder and self-reported physical symptoms in Persian Gulf War veterans. OCLC   772409021.
  27. Kemp, Stephen F. (1995-09-11). "Anaphylaxis: A Review of 266 Cases". Archives of Internal Medicine. 155 (16): 1749–54. doi:10.1001/archinte.1995.00430160077008. ISSN   0003-9926. PMID   7654108.
  28. Warner, J.O. (2002). "How dangerous is food allergy in childhood?". Pediatric Allergy and Immunology. 13 (3): 149–150. doi:10.1034/j.1399-3038.2002.00059.x. ISSN   0905-6157. PMID   12144634. S2CID   10701629.
  29. ScienceDirect (Service en ligne). The journal of pediatrics. OCLC   798778572.
  30. Oki, T.; Yoshimoto, A.; Sato, S.; Takamatsu, A. (1975-12-18). "Purine nucleotide pyrophosphotransferase from Streptomyces morookaensis, capable of synthesizing pppApp and pppGpp". Biochimica et Biophysica Acta (BBA) - Enzymology. 410 (2): 262–272. doi:10.1016/0005-2744(75)90228-4. ISSN   0006-3002. PMID   1088.
  31. Boyd, George K. (1989-07-01). "Fatal Nut Anaphylaxis in a 16-Year-Old Male: Case Report". Allergy and Asthma Proceedings. 10 (4): 255–257. doi:10.2500/108854189778959966. ISSN   1088-5412. PMID   2792751.
  32. Taylor, Stephen L.; Bush, Robert K.; Busse, William W. (1986-11-01). "Avoidance Diets—How Selective Should We Be?". Allergy and Asthma Proceedings. 7 (6): 527–532. doi:10.2500/108854186779045502. ISSN   1088-5412.
  33. Dawson, R. M. (1975). "The reaction of choline and 3,3-dimethyl-1-butanol with the acetylenzyme from acetylcholinesterase". Journal of Neurochemistry. 25 (6): 783–787. doi:10.1111/j.1471-4159.1975.tb04408.x. ISSN   0022-3042. PMID   1471. S2CID   45389128.
  34. Prado, M.; Ortea, I.; Vial, S.; Rivas, J.; Calo-Mata, P.; Barros-Velázquez, J. (2016-11-17). "Advanced DNA- and Protein-based Methods for the Detection and Investigation of Food Allergens". Critical Reviews in Food Science and Nutrition. 56 (15): 2511–2542. doi:10.1080/10408398.2013.873767. ISSN   1040-8398. PMID   25848852. S2CID   21405610.
  35. Brežná, B.; Hudecová, L.; Kuchta, T. (2006). "A novel real-time polymerase chain reaction (PCR) method for the detection of walnuts in food". European Food Research and Technology. 223 (3): 373–377. doi:10.1007/s00217-005-0214-8. ISSN   1438-2377. S2CID   186222380.
  36. Yano, Takeo; Sakai, Yumiko; Uchida, Kohji; Nakao, Yoshiki; Ishihata, Kimie; Nakano, Shigeru; Yamada, Toshihiro; Sakai, Shinobu; Urisu, Atsuo (2007-07-23). "Detection of Walnut Residues in Processed Foods by Polymerase Chain Reaction". Bioscience, Biotechnology, and Biochemistry. 71 (7): 1793–1796. doi: 10.1271/bbb.70118 . ISSN   0916-8451. PMID   17617706.
  37. 1 2 3 4 5 Doi, Hirotoshi; Touhata, Yuki; Shibata, Haruki; Sakai, Shinobu; Urisu, Atsuo; Akiyama, Hiroshi; Teshima, Reiko (2008-09-10). "Reliable Enzyme-Linked Immunosorbent Assay for the Determination of Walnut Proteins in Processed Foods". Journal of Agricultural and Food Chemistry. 56 (17): 7625–7630. doi:10.1021/jf801550h. ISSN   0021-8561. PMID   18681443.
  38. 1 2 Savoye, F.; Feng, P.; Rozand, C.; Bouvier, M.; Gleizal, A.; Thevenot, D. (2011). "Comparative Evaluation of a Phage Protein Ligand Assay with Real-Time PCR and a Reference Method for the Detection of Escherichia coli O157:H7 in Raw Ground Beef and Trimmings". Journal of Food Protection. 74 (1): 6–12. doi: 10.4315/0362-028X.JFP-10-271 . ISSN   0362-028X. PMID   21219756.
  39. Kunze, H.; Bohn, E.; Bahrke, G. (1975). "Effects of psychotropic drugs on prostaglandin biosynthesis in vitro". The Journal of Pharmacy and Pharmacology. 27 (11): 880–881. doi:10.1111/j.2042-7158.1975.tb10239.x. ISSN   0022-3573. PMID   1505. S2CID   20327465.
  40. Anderson, Neil; Suliman, Ibnauf; Bandaletova, Tatiana; Obichere, Austin; Lywood, Rupert; Loktionov, Alexandre (2011). "Protein biomarkers in exfoliated cells collected from the human rectal mucosa: implications for colorectal disease detection and monitoring". International Journal of Colorectal Disease. 26 (10): 1287–1297. doi:10.1007/s00384-011-1263-z. ISSN   0179-1958. PMID   21698353. S2CID   24118612.
  41. Parro, Víctor; Rivas, Luis A.; Gómez-Elvira, Javier (2008). "Protein Microarrays-Based Strategies for Life Detection in Astrobiology". Space Science Reviews. 135 (1–4): 293–311. Bibcode:2008SSRv..135..293P. doi:10.1007/s11214-007-9276-1. ISSN   0038-6308. S2CID   122119527.