[Selection and identification of completely humanized anti-IL-4 single-chain antibodies].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
; 31(12): 1698-701, 2015 Dec.
Article
em Zh
| MEDLINE
| ID: mdl-26648307
ABSTRACT
OBJECTIVE:
To express human interleukin 4 (IL-4), select completely humanized anti-IL-4 single-chain antibodies (scFvs) from the completely humanized scFv library and identify their specificity.METHODS:
With IPTG-induced pET102/IL-4/BL21, human IL-4 was synthesized, and then purified and identified. Completely humanized scFvs against IL-4 were expressed from the completely humanized antibody phage library, and positive clones were selected with human IL-4 as antigen. The positive clones were tested by PCR, enzyme cleavage and sequencing. The specificity of the completely humanized anti-IL-4 scFvs was determined by dot blot hybridization.RESULTS:
The relative molecular mass (Mr) of IL-4 fusion protein was about 27 000. With IL-4 as antigen, we obtained the positive clones expressing anti-IL-4 antibodies. PCR showed amplified target bands of about 1000 bp. Enzyme cleavage analysis revealed that fingerprint maps of the selected clones were diverse except 2 clones. Among 4 clones expressing scFvs that had different fingerprint maps and strong binding to IL-4, sequencing showed the sequence of 3 was correct. Then, dot blot hybridization demonstrated that the scFvs expressed by the 3 clones were specific.CONCLUSION:
Completely humanized scFvs against IL-4 has been obtained successfully.
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Base de dados:
MEDLINE
Assunto principal:
Interleucina-4
/
Anticorpos de Cadeia Única
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Humans
Idioma:
Zh
Revista:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
Assunto da revista:
ALERGIA E IMUNOLOGIA
Ano de publicação:
2015
Tipo de documento:
Article
País de afiliação:
China