[Monoclonal Antibody Against N Terminal 439-451 Epitopes of Anti-Mullerian Hormone and Its Properties].
Sichuan Da Xue Xue Bao Yi Xue Ban
; 51(4): 494-498, 2020 Jul.
Article
em Zh
| MEDLINE
| ID: mdl-32691556
ABSTRACT
OBJECTIVE:
To prepare the specific monoclonal antibody against the N-terminal specific epitope peptide of anti-mullerian hormone (AMH) and to identify its specificity.METHODS:
Using bioinformatics analysis software to predict the specific peptide fragment of AMH. Then synthesized four antigenic epitope peptide segments of mature N-terminal region of AMH as the screening target antigen. Synthesized AMH wholegene.Using the prokaryotic expression system to abtain recombinant AMH protein. Immunized BALB/c mice with the recombinant AMH, and prepared mouse spleen cells for fusing with SP/20 cells. Preparation of AMH monoclonal antibody by hybridoma technology. The monoclonal antibodies against AMH were screened by using four N-terminal epitope peptides (1 439-451 RGRDPRGPGRAQ, 2 273-285 PPRPSAELEESPP, 3 42-54 DLDWPPGSPQEPL, 4 494-506 WPQSDRNPRYGNH) as antigens, and indirect ELISA and Western blot were used to identify the antigen binding characteristics of the selected monoclonal antibodies.RESULTS:
Two hybridoma cell lines with stable anti-AMH-1 and anti-AMH-2 antibody activities were screened. The two antibodies were named anti-AMH-1 and anti-AMH-2 respectively. The antibody titers were 1â¶12 000 and 1â¶1 600 after purification. Western blot confirmed that the two McAbs recognized different antigens. Anti-AMH-1 could not only recognize the N-terminal 439-451 epitope peptide of AMH, but also recognize the amino acid sequence of recombinant AMH, as well as the ovarian tissue. Anti-AMH-2 could recognize recombinant AMH and ovarian tissue.CONCLUSION:
Two monoclonal antibodies against N-terminal specific epitopes of human AMH were successfully constructed.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Hormônio Antimülleriano
/
Anticorpos Monoclonais
/
Epitopos
Tipo de estudo:
Prognostic_studies
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Sichuan Da Xue Xue Bao Yi Xue Ban
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
China