Production, characterization, and applications of mouse monoclonal antibodies against gonadotropin, somatolactin, and prolactin from common carp (Cyprinus carpio).
The gonadotropin alpha subunit (cGTH alpha), gonadotropin II beta subunit (cGTHII beta), somatolactin (cSL), and prolactin (cPRL) were isolated from the pituitaries of common carps, purified by traditional chromatographic analysis, identified by mass-chromatographic analysis, and used as immunogens in the B-lymphocyte hybridoma technique. Totally, 7, 11, 17, and 8 hybridoma cell lines were established, which were able to stably secrete monoclonal antibodies (mAbs) against cGTH alpha, cGTHII beta, cSL, and cPRL, and designated as FMU-cGTH alpha 1-7, FMU-cGTHII beta 1-11, FMU-cSL 1-17, and FMU-cPRL 1-8, respectively. The isotype, titer, and specificity were identified by enzyme-linked immunosorbent assay (ELISA), Western blot, and immunohistochemical staining, respectively, and application of these mAbs in the aforementioned tests has been proved. Furthermore, sensitive sandwich-ELISA systems for quantitative detection of the hormones mentioned above were also developed.
Department of Immunology, The Fourth Military Medical University, Xi'an 710032, China.
Zhuwei
Z
Xu
Qi
Q
Li
Yun
Y
Wang
Chaojun
C
Song
Tao
T
Zhang
Lihua
L
Chen
Jianguo
J
Ji
Angang
A
Yang
Zuoyan
Z
Zhu
Wei
W
Hu
Boquan
B
Jin
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology*; D000917:Antibody Formation; D002347:Carps / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D005260:Female; D029941:Fish Proteins / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D006023:Glycoproteins / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D006062:Gonadotropins / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D007150:Immunohistochemistry / Q000592:standards; D051379:Mice; D008807:Mice, Inbred BALB C; D010907:Pituitary Hormones / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D011388:Prolactin / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D012015:Reference Standards
D000911:Antibodies, Monoclonal; D029941:Fish Proteins; D006023:Glycoproteins; D006062:Gonadotropins; D010907:Pituitary Hormones; C064448:somatolactin protein, fish; D011388:Prolactin