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Expression of an antibody-enzyme complex by the L-chain fusion method

Paper ID Volume ID Publish Year Pages File Format Full-Text
20844 43194 2013 5 PDF Available
Title
Expression of an antibody-enzyme complex by the L-chain fusion method
Abstract

In this report, we describe a novel method for directly preparing enzyme-labeled antibodies harvested from IgM-producing hybridoma cells. We constructed expression vectors for antibody light (L) chain-enzyme fusion proteins by linking either the genes for the murine lambda L chain or its constant region (CL) with one of two proteins, either the secreted placental alkaline phosphatase or Gaussia luciferase (Gluc). When the vectors were transfected into anti-NP (4-hydroxy-3-nitrophacetyl) IgM-producing myeloma cells, secretion of the IgM-enzyme complex from the gene-transfected cells was confirmed by a direct enzyme-linked immunosorbent assay with an immobilized antigen. Furthermore, when human hybridoma HF10B4, a cell line that produces anti-human lung cancer IgM, was transfected with the vector containing L-Gluc, a significantly stronger signal was obtained for the human lung carcinoma SBC-1 cells than for cervical HeLa cells. Because successful production of an active IgM-enzyme complex containing a heterologous L chain-enzyme fusion was observed, the L-chain fusion method will be a generally applicable method for preparing various IgM-enzyme complexes.

► A method is proposed for preparing enzyme-labeled antibodies from hybridoma cells. ► Vectors for antibody light (L) chain-enzyme fusion proteins were transfected. ► Secretion of functional IgM–enzyme complex from the transfected cells was confirmed. ► The method was successfully applied to the detection of human lung carcinoma cells.

Keywords
4-Hydroxy-3-nitrophenacetyl; Human lung carcinoma; Enzyme-linked immunosorbent assay; Fusion protein; Secreted alkaline phosphatase; Gaussia luciferaseBSA, bovine serum albumin; CL, constant region fragment of L chain; ELISA, enzyme-linked immunosorbent a
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 116, Issue 1, July 2013, Pages 17–21
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