绵羊肺腺瘤病毒囊膜蛋白单克隆抗体的制备及双抗体夹心ELISA方法的初步建立

doi:10.11843/j.issn.0366-6964.2015.05.016

Abstract

Abstract:

In order to prepare monoclonal antibody against the cytoplasmic tail of the envelope protein of Jaagsiekte sheep retrovirus（JSRV），the amino acid sequence of the JSRV envelope protein was analyzed by bioinformatics to synthetize polypeptide.Then the polypeptide was connected with Keyhole Limpet hemocyanin（CT-KLH） or bovine serum albumin（CT-BSA），respectively.Polypeptide CT-KLH was immunized to the BALB/c mice as antigen，and the spleen cells of immunized mice were fused with SP2/0 myeloma cells.An indirect ELISA was developed to screen positive antibody producing cells using the CT protein.A hybridoma stably secreting MAb designated as 8E5 was obtained against CT protein.The monoclonal antibody belonged to IgG2a/κ.The titers of supernatant and ascites were 6.4×10³ and 1×10⁵ as detected by ELISA，respectively.The results of western blot and immunohistochemistry showed that this hybridoma secreted MAb could react specifically with the JSRV.The results suggested that the MAb-8E5 is important diagnostic reagents for detection of ovine pulmonary adenocarcinoma (OPA).A sandwich ELISA was established by using 8E5 secreted monoclonal antibody.The results indicated that the ELISA possessed good specificity and repeatability，and higher sensitivity.The monoclonal antibody could be used in diagnosis for OPA and further research of envelope protein function.

CLC Number:

S852.659.3

LIU Yue，ZHANG Yu-fei，SUN Xiao-lin，LIU Shu-ying. Preparation of the Monoclonal Antibody against Envelope Protein of JSRV and Development of Double Antibody Sandwich ELISA for JSRV[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(5): 800-807.

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Preparation of the Monoclonal Antibody against Envelope Protein of JSRV and Development of Double Antibody Sandwich ELISA for JSRV

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