Generation and Characterization of Monoclonal Antibodies against Human Interferon-lambda1
Hong, SH; Kim, JS; Park, S
Immune network, 8(1):7-12, 2008
Background:Members belonging to the interferon-lambda (IFN-?) family exert protective action against viral infection; however, the mechanisms of their action have remained elusive. To study IFN-? biology, such as endocytosis of IFN-?, we produced monoclonal antibodies (Abs) against human IFN-? and examined their usefulness.
Methods:We purified recombinant human IFN-?1 expressed in Escherichia coli by using affinity columns. Then, we generated hybridoma cells by fusing myeloma cells with splenocytes from IFN-?1- immunized mice. For evaluating the neutralizing activity of the monoclonal Abs against IFN-?1, we performed RT-PCR for the MxA transcript. In order to study the binding activity of IFN-? and the monoclonal Ab complex on HepG2 cells, we labeled the monoclonal Ab with rhodamine and determined the fluorescence intensity.
Results:Four hybridoma clones secreting Abs specific to IFN-?1 were generated and designated as HL1, HL2, HL3, and HL4. All the Abs reacted with IFN-?1 in the denatured form as well as in the native form. Abs produced by HL1, HL3, and HL4 did not neutralize the induction of the MxA gene by IFN-?1. We also demonstrated the binding of the HL1 monoclonal anbitody and IFN-? complex on HepG2 cells.
Conclusion:Monoclonal Abs against IFN-?1 were produced. These Abs can be used to study the cellular binding and internalization of IFN-?.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.
Total Visit :5,294,365
Total Download :2,237,075
Today View :3,920
Ajou University Medical Information & Media Center 164 Worldcup-ro Yeongtong-gu Suwon 16499 Korea / TEL : 031-219-5312 / FAX : 031-219-5314 Copyright (c) Ajou University Medical Information & Media Center All Rights Reserved. AJOU Open Repository는 국립중앙도서관 OAK 보급사업으로 구축되었습니다.