Application of avidin-biotin technology for the characterization of a model hapten-protein conjugate

被引：3

作者：

Dotsikas, Y ^{[1
]}

Loukas, YL ^{[1
]}

机构：

[1] Univ Athens, Sch Pharm, Dept Pharmaceut Chem, GR-15771 Athens, Greece

来源：

JOURNAL OF IMMUNOASSAY & IMMUNOCHEMISTRY | 2005年 / 26卷 / 04期

关键词：

hapten-BSA conjugate; electrospray mass spectrometry; HABA; biotin;

D O I：

10.1080/15321810500220845

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A simple method was developed for the rapid characterization of the covalent binding of haptens to proteins such as enzymes, bovine serum albumin (BSA), and other carrier-proteins and antibodies. In the present study, a commercially available fentanyl-BSA conjugate was characterized by a 4'-hydroxyazobenzene-2-carboxylic acid (HABA) dye assay that followed a biotinylation reaction. This protocol allowed the indirect observation of the average hapten number per BSA molecule. Such measurement is useful for optimizing reaction conditions to yield a more precisely defined product for immunological applications. The obtained result was within the limits suggested by the manufacturer of the conjugate.

引用

页码：285 / 293

页数：9

共 25 条

[1]

BARBARAKIS MS, 1991, CLIN CHEM, V37, P87

[2]

COLE RB, 1997, ELECTROSPRAY IONIZAT, V55

[3] Employment of 4-(1-imidazolyl)phenol as a luminol signal enhancer in a competitive-type chemiluminescence immunoassay and its comparison with the conventional antigen-horseradish peroxidase conjugate-based assay [J].

Dotsikas, Y ;

Loukas, YL .

ANALYTICA CHIMICA ACTA, 2004, 509 (01) :103-109

[4] Kinetic degradation study of insulin complexed with methyl-beta cyclodextrin.: Confirmation of complexation with electrospray mass spectrometry and 1H NMR [J].

Dotsikas, Y ;

Loukas, YL .

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, 2002, 29 (03) :487-494

[5] Determination of umbilical cord and maternal plasma concentrations of fentanyl by using novel spectrophotometric and chemiluminescence enzyme immunoassays [J].

Dotsikas, Y ;

Loukas, YL ;

Siafaka, I .

ANALYTICA CHIMICA ACTA, 2002, 459 (02) :177-185

[6] A GENERAL-METHOD FOR DEVELOPING IMMUNOASSAYS TO CHLOROACETANILIDE HERBICIDES [J].

FENG, PCC ;

HORTON, SR ;

SHARP, CR .

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1992, 40 (02) :211-214

[7] STUDIES ON THE BIOTIN-BINDING SITE OF AVIDIN - LYSINE RESIDUES INVOLVED IN THE ACTIVE-SITE [J].

GITLIN, G ;

BAYER, EA ;

WILCHEK, M .

BIOCHEMICAL JOURNAL, 1987, 242 (03) :923-926

[8]

GOSSLING JP, 2000, IMMUNOASSAYS PRACTIC, V172

[9] DETERMINATION OF FREE AMINO GROUPS IN PROTEINS BY TRINITROBENZENESULFONIC ACID [J].

HABEEB, AFS .

ANALYTICAL BIOCHEMISTRY, 1966, 14 (03) :328-&

[10]

HENDERSON GL, 1975, J PHARMACOL EXP THER, V192, P489

← 1 2 3 →