Preparation and Characteristic of Monoclonal Antibody against Phytase

doi:10.3969/j.issn.1000-7091.2012.05.009

Abstract

Abstract: The hybridoma lines that secrete phytase monoclonal antibody(mAb) were established through cell fusion.Phytase mAb was prepared by inducing ascites in mice.The immunological characteristics of the mAb were identified.The result indicated that two hybridomas were selected.The subtype of mAb generated by 5C3 hybridoma was IgG1,the titer was 1 ∶ 1.02×106,IC50 was 100.07 ng/mL and Kaff was 6.70×109 L/mol.Phytase mAb IC50 to normal,coated and concentrated phytase was 130.43,126.16,125.33 ng/mL,and cross-reactivity was 76.72%,79.32% and 79.84%,respectively.Phytase mAb IC50 to inactivated(boiled for 5 min) wheat bran phytase,normal,coated and concentrated phytase was 4 205.45,4 015.86,3 593.19,3 610.40 ng/mL,and the cross-reactivity was 2.38%,2.49%,2.78% and 2.77%,respectively.In conclusion,hybridoma lines that secrete phytase mAb were established,phytase mAb with high affinity and specificity and sensitive was prepared.The phytase mAb may lay a foundation for the subsequent ELISA kit and strip development.

Key words: Phytase, Hybridoma mAb, ELISA

CLC Number:

S816

HU Xiao-fei, WEI Feng-xian, LI Qing-mei, ZHI Ai-min, WANG Fang-yu, SUN Ya-ning, CHAI Shu-jun, FU Xiao-he. Preparation and Characteristic of Monoclonal Antibody against Phytase[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(5): 44-48. doi: 10.3969/j.issn.1000-7091.2012.05.009.

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