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

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

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.
LanguageEnglish
Pages1441-1444
Number of pages4
JournalVaccine
Volume13
Issue number15
DOIs
Publication statusPublished - 1995

Fingerprint

Ninhydrin
Aluminum Hydroxide
adjuvants
Subunit Vaccines
peptides
antigens
Liposomes
aluminum hydroxide
Peptides
subunit vaccines
assays
Immunologic Adjuvants
Proteins
proteins
Immunity
Hydrolysis
amino acid composition
Safety
Antigens
Amino Acids

Keywords

  • adjuvants, immunologic
  • aluminum hydroxide
  • liposomes
  • measles vaccine
  • ninhydrin
  • ovalbumin
  • peptides
  • surface-active agents
  • viral proteins

Cite this

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title = "Accurate determination of adjuvant-associated protein or peptide by ninhydrin assay",
abstract = "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.",
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author = "Brewer, {J M} and Roberts, {C W} and Stimson, {W H} and J Alexander",
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Accurate determination of adjuvant-associated protein or peptide by ninhydrin assay. / Brewer, J M; Roberts, C W; Stimson, W H; Alexander, J.

In: Vaccine, Vol. 13, No. 15, 1995, p. 1441-1444.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Accurate determination of adjuvant-associated protein or peptide by ninhydrin assay

AU - Brewer, J M

AU - Roberts, C W

AU - Stimson, W H

AU - Alexander, J

PY - 1995

Y1 - 1995

N2 - 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.

AB - 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.

KW - adjuvants, immunologic

KW - aluminum hydroxide

KW - liposomes

KW - measles vaccine

KW - ninhydrin

KW - ovalbumin

KW - peptides

KW - surface-active agents

KW - viral proteins

U2 - 10.1016/0264-410X(95)00065-9

DO - 10.1016/0264-410X(95)00065-9

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T2 - Vaccine

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