Generation and Characterization of Monoclonal Antibodies against Human Interferon-lambda1
Immune Network
;
: 7-12, 2008.
Article
in English
| WPRIM
| ID: wpr-142420
ABSTRACT
BACKGROUND:
Members belonging to the interferon-lambda (IFN-lambda) family exert protective action against viral infection; however, the mechanisms of their action have remained elusive. To study IFN-lambda biology, such as endocytosis of IFN-lambda, we produced monoclonal antibodies (Abs) against human IFN-lambda and examined their usefulness.METHODS:
We purified recombinant human IFN-lambda1 expressed in Escherichia coli by using affinity columns. Then, we generated hybridoma cells by fusing myeloma cells with splenocytes from IFN-lambda1- immunized mice. For evaluating the neutralizing activity of the monoclonal Abs against IFN-lambda1, we performed RT-PCR for the MxA transcript. In order to study the binding activity of IFN-lambda 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-lambda1 were generated and designated as HL1, HL2, HL3, and HL4. All the Abs reacted with IFN-lambda1 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-lambda1. We also demonstrated the binding of the HL1 monoclonal anbitody and IFN-lambda complex on HepG2 cells.CONCLUSION:
Monoclonal Abs against IFN-lambda1 were produced. These Abs can be used to study the cellular binding and internalization of IFN-lambda.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Rhodamines
/
Biology
/
Clone Cells
/
Endocytosis
/
Escherichia coli
/
Hep G2 Cells
/
Fluorescence
/
Hybridomas
/
Antibodies, Monoclonal
Limits:
Animals
/
Humans
Language:
English
Journal:
Immune Network
Year:
2008
Type:
Article
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