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Generation and Characterization of Monoclonal Antibodies against Human Interferon-lambda1

Authors
Hong, SH; Kim, JS; Park, S
Citation
Immune network, 8(1):7-12, 2008
Journal Title
Immune network
ISSN
1598-26292092-6685
Abstract
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-?.
Keywords
Interleukin 29 proteinmonoclonal antibody
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
AJOU Authors
박, 선
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