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Enzyme linked immunosorbent assay using alkaline phosphatase conjugated with streptococcal protein G

Nilson, B LU ; Björck, L LU and Akerström, B LU (1988) In Journal of immunoassay 9(2). p.25-207
Abstract

Protein G, an IgG-binding protein, purified from the surface of group G streptococci, was coupled to alkaline phosphatase. The conjugate was used for detection of polyclonal goat and rabbit antibodies and monoclonal mouse IgG1, IgG2a and IgG2b in an enzyme-linked immunosorbent assay. A two-step coupling procedure was used, in which glutaraldehyde was allowed to react with the enzyme, excess glutaraldehyde was then removed by dialysis, and finally protein G added to the glutaraldehyde-activated and polymerized alkaline phosphatase. The activity and yield of the conjugates were then tested in an enzyme-linked immunosorbent assay. Coupling of 25 micrograms protein G to 5 mg alkaline phosphatase gave a conjugate which could be used for more... (More)

Protein G, an IgG-binding protein, purified from the surface of group G streptococci, was coupled to alkaline phosphatase. The conjugate was used for detection of polyclonal goat and rabbit antibodies and monoclonal mouse IgG1, IgG2a and IgG2b in an enzyme-linked immunosorbent assay. A two-step coupling procedure was used, in which glutaraldehyde was allowed to react with the enzyme, excess glutaraldehyde was then removed by dialysis, and finally protein G added to the glutaraldehyde-activated and polymerized alkaline phosphatase. The activity and yield of the conjugates were then tested in an enzyme-linked immunosorbent assay. Coupling of 25 micrograms protein G to 5 mg alkaline phosphatase gave a conjugate which could be used for more than 10,000 determinations with maximal antibody binding giving an absorbance of 2.0. Under these conditions, there was no need for separation of the reactants before using the protein G-alkaline phosphatase complex.

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organization
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type
Contribution to journal
publication status
published
subject
keywords
Alkaline Phosphatase, Antibodies, Monoclonal, Bacterial Proteins, Chromatography, Gel, Enzyme-Linked Immunosorbent Assay, Immunoglobulin G, Temperature, Time Factors, Journal Article, Research Support, Non-U.S. Gov't
in
Journal of immunoassay
volume
9
issue
2
pages
25 - 207
external identifiers
  • scopus:0023895639
ISSN
0197-1522
DOI
10.1080/15321818808057041
language
English
LU publication?
yes
id
914ccc37-7f85-45f1-aa6f-d60851c66478
date added to LUP
2018-05-26 14:10:38
date last changed
2018-06-03 05:07:03
@article{914ccc37-7f85-45f1-aa6f-d60851c66478,
  abstract     = {<p>Protein G, an IgG-binding protein, purified from the surface of group G streptococci, was coupled to alkaline phosphatase. The conjugate was used for detection of polyclonal goat and rabbit antibodies and monoclonal mouse IgG1, IgG2a and IgG2b in an enzyme-linked immunosorbent assay. A two-step coupling procedure was used, in which glutaraldehyde was allowed to react with the enzyme, excess glutaraldehyde was then removed by dialysis, and finally protein G added to the glutaraldehyde-activated and polymerized alkaline phosphatase. The activity and yield of the conjugates were then tested in an enzyme-linked immunosorbent assay. Coupling of 25 micrograms protein G to 5 mg alkaline phosphatase gave a conjugate which could be used for more than 10,000 determinations with maximal antibody binding giving an absorbance of 2.0. Under these conditions, there was no need for separation of the reactants before using the protein G-alkaline phosphatase complex.</p>},
  author       = {Nilson, B and Björck, L and Akerström, B},
  issn         = {0197-1522},
  keyword      = {Alkaline Phosphatase,Antibodies, Monoclonal,Bacterial Proteins,Chromatography, Gel,Enzyme-Linked Immunosorbent Assay,Immunoglobulin G,Temperature,Time Factors,Journal Article,Research Support, Non-U.S. Gov't},
  language     = {eng},
  number       = {2},
  pages        = {25--207},
  series       = {Journal of immunoassay},
  title        = {Enzyme linked immunosorbent assay using alkaline phosphatase conjugated with streptococcal protein G},
  url          = {http://dx.doi.org/10.1080/15321818808057041},
  volume       = {9},
  year         = {1988},
}