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PMID: 6351851 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A monoclonal antibody to kidney endopeptidase-24.11. Its application in immunoadsorbent purification of the enzyme and immunofluorescent microscopy of kidney and intestine.

The Biochemical journal ·Vol. 214 ·No. 2 ·1983-08-15 ·Pages 377-86

Gee NS, Matsas R, Kenny AJ

Abstract

Hybridoma methodology has been used to produce a monoclonal antibody, GK 7C2, that binds specifically to microvillar endopeptidase-24.11 (EC 3.4.24.11). The antibody (an immunoglobulin G) was generated by fusion of mouse plasmacytoma cells with splenocytes from a Balb/c mouse immunized with pig kidney microvillar membranes. The identity of the antigen recognized by GK 7C2 was established by immuno-precipitation from detergent-solubilized pig kidney microvilli. The protein had an apparent Mr of 90 000 and contained endopeptidase activity sensitive to phosphoramidon. The identity was confirmed by immunoadsorbent purification of endopeptidase-24.11 by a column to which GK 7C2 had been attached. The endopeptidase, purified in a yield of 40%, was electrophoretically homogeneous and of specific activity comparable with that purified by other means. Fluorescence microscopy established that GK 7C2 bound specifically to the luminal membranes of kidney tubules and the intestinal mucosa. Thus endopeptidase-24.11 is located in the brush-border membranes of both cell types.

MeSH Terms

Animals Antibodies, Monoclonal/immunology Cells, Cultured Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique Immunoglobulin G/immunology Immunosorbent Techniques Jejunum/enzymology Kidney/enzymology Mice Mice, Inbred BALB C Microscopy, Fluorescence Microscopy, Phase-Contrast Microvilli/enzymology Swine

Chemicals

Antibodies, Monoclonal Immunoglobulin G

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Gee N S

Matsas R

Kenny A J

References (21)

21 references, click to expand

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Article Info

Journal

The Biochemical journal

Abbr.

Biochem J

ISSN

0264-6021

DOI

10.1042/bj2140377

Published

1983-08-15

Pages

377-86

Language

English

Region

England

NLM ID

2984726R

PMCID

PMC1152258

Subset

External Links

PubMed source DOI: 10.1042/bj2140377 PMC Full Text View journal details

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A monoclonal antibody to kidney endopeptidase-24.11. Its application in immunoadsorbent purification of the enzyme and immunofluorescent microscopy of kidney and intestine.

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