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Analysis of cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulin

Paper ID Volume ID Publish Year Pages File Format Full-Text
20988 43200 2013 7 PDF Available
Title
Analysis of cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulin
Abstract

Although various in vitro immunization methods to generate antigen-specific antibodies have been described, a highly effective method that can generate high-affinity immunoglobulins has not yet been reported. Herein, we analyzed a cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulins. We identified a combination of T cell-dependent stimuli (IL-4, IL-5, anti-CD38 and anti-CD40 antibodies) plus lipopolysaccharides (LPS) that stimulates antigen-exposed splenocytes in vitro, followed by induction of the cells phenotypically equivalent to germinal center B cells. We also observed that LPS induced high expression levels of mRNA for activation-induced cytidine deaminase. We stimulated antigen-exposed splenocytes, followed by the accumulation of mutations in immunoglobulin genes. From the immunized splenocytes, hybridoma clones secreting antigen-specific immunoglobulins were obtained.

Keywords
In vitro immunization; Activation-induced cytidine deaminase; Germinal center; Somatic hypermutation; T cell dependent stimuli
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Analysis of cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulin
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 115, Issue 3, March 2013, Pages 339–345
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us