Accurate determination of adjuvant-associated protein or peptide by ninhydrin assay

Modern peptide and subunit vaccines are increasingly having to rely on the use of immunological adjuvants to achieve effective immunity. However, the only adjuvant currently approved for use in humans is aluminium hydroxide, although many adjuvants are currently under preclinical development. Determ...

Full description

Saved in:
Bibliographic Details
Published in:Vaccine 1995, Vol.13 (15), p.1441-1444
Main Authors: Brewer, J.M., Roberts, C.W., Stimson, W.H., Alexander, J.
Format: Article
Language:English
Subjects:
Adjuvant
Adjuvants, Immunologic - analysis
alhydrogel
Aluminum Hydroxide - analysis
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Liposomes - analysis
Measles Vaccine - analysis
Molecular immunology
Ninhydrin
non-ionic surfactant vesicle
Ovalbumin - analysis
Peptides - analysis
Peptides - immunology
protein assay
Surface-Active Agents - analysis
Techniques
Viral Proteins - analysis
Viral Proteins - immunology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Modern peptide and subunit vaccines are increasingly having to rely on the use of immunological adjuvants to achieve effective immunity. However, the only adjuvant currently approved for use in humans is aluminium hydroxide, although many adjuvants are currently under preclinical development. Determining immunogen concentration in the presence of adjuvants such as aluminium hydroxide gel, liposomes or NISV has proved to be problematic. One approach has been to use radiolabelled antigens to extrapolate concentration to a preparation using native immunogen. However, the use of a colorimetric assay would allow greater flexibility in terms of immunogen used and would reduce costs and remove safety problems. Of the colorimetric methods we have examined thus far, only the manual ninhydrin assay has produced consistent results with detection of microgram quantities of protein or peptide in the presence of NISV or Alhydrogel, but not liposomes. As the assay relies on the detection of free amino groups after protein hydrolysis, peptides as well as proteins may be effectively determined irrespective of amino acid composition, a considerable advantage over other colorimetric assay systems.
ISSN:0264-410X
1873-2518
DOI:10.1016/0264-410X(95)00065-9