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KMID : 0923620080080010007
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Immune Network
2008 Volume.8 No. 1 p.7 ~ p.12
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Generation and Characterization of Monoclonal Antibodies against Human Interferon-lambda1
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Hong Seung-Ho
Kim Jung-Sik
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Abstract
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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-?. [Immune Network 2008;8(1):7-12]
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KEYWORD
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Interleukin 29 protein, monoclonal antibody
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