The optimization of ELISA for methamphetamine determination

The effect of immunogen, tracer and antibody purification method on the sensitivity

Jeongeun Choi, Myung Ja Choi, Choonmi Kim, Young Shik Cho, Jaeho Chin, Young Ah Jo

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

To obtain more sensitive immunoassay for methamphetamine (MA) determination, the optimum condition of enzyme-linked immunosorbent assay (ELISA) was investigated in regard to immunogens, antibody purification methods and coating tracers. Activated MA, N-(4-aminobutyl)methamphetamine (4-ABMA), was conjugated with bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) and used as immunogen. The antibodies were purified by protein G chromatography or various immunoaffinity chromatography-linked MA-protein ligands, such as MA-BSA, MA-KLH or MA-ovalbumin (OVA). Each purified antibody was characterized by means of sensitivity and cross-reactivity using the three MA-protein coating tracers, MA-BSA, MA-KLH and MA-OVA. The best sensitivity of each antibody was acquired with the MA-OVA tracer although the tracer concentration and the antibody titer level at optimum condition were varied. The antibody with high titer level did not always yield good sensitivity. At optimum condition, immunoaffinity chromatography-purified antibodies were better for sensitivity and for specificity than protein G-purified antibodies. The cross-reactivity of the ] purified antibodies seemed to be affected by immunogen structure and showed somewhat different patterns according to the immunoaffinity ligand utilized. These data show that the antibody purification method as well as choice of coating tracer and immunogen is essential for the sensitivity and specificity of EIA; the optimum condition for assay should be discovered using various methods and combinations.

Original languageEnglish (US)
Pages (from-to)46-52
Number of pages7
JournalArchives of Pharmacal Research
Volume20
Issue number1
StatePublished - Feb 1997

Fingerprint

Immunosorbents
Methamphetamine
Purification
Assays
Enzyme-Linked Immunosorbent Assay
Antibodies
Enzymes
Ovalbumin
Bovine Serum Albumin
Chromatography
Coatings
Proteins
Ligands
Sensitivity and Specificity
Immunoassay

Keywords

  • Antibody purification method
  • Coating tracer
  • ELISA
  • Methamphetamine
  • Optimization

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Organic Chemistry
  • Drug Discovery
  • Pharmacology

Cite this

The optimization of ELISA for methamphetamine determination : The effect of immunogen, tracer and antibody purification method on the sensitivity. / Choi, Jeongeun; Choi, Myung Ja; Kim, Choonmi; Cho, Young Shik; Chin, Jaeho; Jo, Young Ah.

In: Archives of Pharmacal Research, Vol. 20, No. 1, 02.1997, p. 46-52.

Research output: Contribution to journalArticle

Choi, Jeongeun ; Choi, Myung Ja ; Kim, Choonmi ; Cho, Young Shik ; Chin, Jaeho ; Jo, Young Ah. / The optimization of ELISA for methamphetamine determination : The effect of immunogen, tracer and antibody purification method on the sensitivity. In: Archives of Pharmacal Research. 1997 ; Vol. 20, No. 1. pp. 46-52.
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