Ming PANG, Chen-Xiang ZUO, Ming-Jie LIU, Saad REHMAN, Qin-Lei FAN, Ping LU, De-Kun CHEN, Wen-Tao MA

Development of a Monoclonal Antibody to Detect αs1-casein in the Milk of Healthy and Mastitis-Affcted Goats

This study aimed to evaluate the expression level of caseins during mastitis of goats. Whole goat caseins were used as primary antigens for mouse immunization. A monoclonal antibody (mAb) named 5B with high specificity to goat αs1-casein was developed. Further results showed that mAb 5B can successfully be applied to western blot and ELISA. In addition, immunoflorescence analysis using this mAb showed increased milk αs1-casein level in mastitis-affcted goats. In conclusion, this study has established an effctive tool to evaluate the expression level of αs1-casein during mastitis development.

Sağlıklı ve Mastitisli Keçi Sütünde αs1-kazeini Belirleyen Monoklonal Antikor Geliştirilmesi

Bu çalışma keçilerde mastitis sırasında kazein ekspresyon seviyesini değerlendirmeyi amaçlamıştır. Farelerde immünizasyon için birincil antijen olarak keçi kazeinleri kullanıldı. Keçi αs1-kazeine yüksek spesifite gösteren ve 5B olarak adlandırılan bir monoklonal antikor (mAb) geliştirildi. Sonuçlar mAb 5B’nin western blot ve ELISA’ya başarıyla uygulanabileceğini gösterdi. Ek olarak, geliştirilen mAb kullanılarak yapılan immünoflresan analizi, mastitisten etkilenen keçilerde süt αs1-kazein seviyesinin arttığını gösterdi. Sonuç olarak, bu çalışma ile mastitis gelişimi sırasında αs1-kazeinin ekspresyon seviyesini değerlendirmek için etkili bir faktör belirlenmiştir.

PDF

___

1. Clark S, Mora Garcia MB: A 100-year review: Advances in goat milk research. J Dairy Sci, 100 (12): 10026-10044, 2017. DOI: 10.3168/jds.2017- 13287

2. Turck D: Cow’s milk and goat’s milk. World Rev Nutr Diet, 108, 56-62, 2013. DOI: 10.1159/000351485

3. Haenlein GFW: Goat milk in human nutrition. Small Ruminant Res, 51 (2): 155-163, 2004. DOI: 10.1016/j.smallrumres.2003.08.010

4. Moreno-Montoro M, Olalla-Herrera M, Rufian-Henares JA, Martinez RG, Miralles B, Bergillos T, Navarro-Alarcon M, Jauregi P: Antioxidant, ACE-inhibitory and antimicrobial activity of fermented goat milk: Activity and physicochemical property relationship of the peptide components. Food Funct, 8, 2783-2791, 2017. DOI: 10.1039/c7fo00666g

5. Liao Y, Weber D, Xu W, Durbin-Johnson BP, Phinney BS, Lonnerdal B: Absolute quantification of human milk caseins and the whey/casein ratio during the first year of lactation. J Proteome Res, 16 (11): 4113-4121, 2017. DOI: 10.1021/acs.jproteome.7b00486

6. Ma W, Wang Y, Gao F, Ning M, Liu A, Li Y, Gao Y, Lu P, Chen D: Development of a monoclonal antibody against bovine α-casein to evaluate functional status of mammary epithelial cells during mastitis. Kafkas Univ Vet Fak Derg, 25 (4): 445-450, 2019. DOI: 10.9775/kvfd.2018.20897

7. Mahlangu P, Maina N, Kagira J: Prevalence, risk factors, and antibiogram of bacteria isolated from milk of goats with subclinical mastitis in Thika East Subcounty, Kenya. J Vet Med, 2018: 3801479, 2018. DOI: 10.1155/2018/3801479

8. Li N, Ma W, Shen Q, Zhang M, Du Z, Wu C, Niu B, Liu W, Hua J: Reconstitution of male germline cell specification from mouse embryonic stem cells using defined factors in vitro. Cell Death Diffr, 2019. DOI: 10.1038/s41418-019-0280-2

9. Schulmeister U, Hochwallner H, Swoboda I, Focke-Tejkl M, Geller B, Nystrand M, Harlin A, Thalhamer J, Scheiblhofer S, Keller W, Niggemann B, Quirce S, Ebner C, Mari A, Pauli G, Herz U, Valenta R, Spitzauer S: Cloning, expression, and mapping of allergenic determinants of alphaS1-casein, a major cow’s milk allergen. J Immunol, 182 (11): 7019-7029, 2009. DOI: 10.4049/jimmunol.0712366

10. Castillo DS, Cassola A: Novel sensitive monoclonal antibody based competitive enzyme-linked immunosorbent assay for the detection of raw and processed bovine beta-casein. PLoS One, 12 (7): e0182447, 2017. DOI: 10.1371/journal.pone.0182447

11. Summer A, Santus E, Casanova L, Joerg H, Rossoni A, Nicoletti C, Donofrio G, Mariani P, Malacarne M: Characterization of a monoclonal antibody for kappa-casein B of cow’s milk. J Dairy Sci, 93 (2): 796-800, 2010. DOI: 10.3168/jds.2009-2636

12. Cakebread J, Hodgkinson A, Wallace O, Callaghan M, Hurford D, Wieliczko R, Harris P, Haigh B: Bovine milk derived skimmed milk powder and whey protein concentrate modulates Citrobacter rodentium shedding in the mouse intestinal tract. PeerJ, 6: e5359, 2018. DOI: 10.7717/ peerj.5359

13. Sicherer SH: Clinical implications of cross-reactive food allergens. J Allergy Clin Immunol, 108 (6): 881-890, 2001. DOI: 10.1067/mai.2001.118515

14. Kobayashi K, Oyama S, Uejyo T, Kuki C, Rahman MM, Kumura H: Underlying mechanisms involved in the decrease of milk secretion during Escherichia coli endotoxin induced mastitis in lactating mice. Vet Res, 44:119, 2013. DOI: 10.1186/1297-9716-44-119