Details
| Original language | English |
|---|---|
| Pages (from-to) | 4027-4033 |
| Number of pages | 7 |
| Journal | Analytical chemistry |
| Volume | 66 |
| Issue number | 22 |
| Publication status | Published - 15 Nov 1994 |
| Externally published | Yes |
Abstract
Antibodies from human sera (h-IgG) were tagged with a fluorescent dye (fluorescein isothiocyanate, FITC) through the affinity reaction of FITC-labeled protein G with the Fc fragment of the antibodies. The complexes were quantified by capillary zone electrophoresis (CZE) within 1 min, i.e., fast enough to prevent their dissociation during the measurement. Conditions for the affinity reaction and the CZE analysis could thus be optimized independently. When an FITC-labeled protein G concentration of 10−6 mol/L was used, h-IgG concentrations between 10−6 and 10−9 mol/L were reproducibly quantified (STD < 2%), using an LIF detector. A correlation coefficient, r2, of 0.9988 was established between the peak height and the IgG concentration. Alternatively, h–IgG containing serum samples and the FITC-labeled protein G were simply injected into the CE capillary in consecutive zones, followed by the application of the electrical field. Within 2 min, the affinity complexes were resolved and the IgG content of the serum quantified (r2 = 0.9986). The injection sequence was of no consequence. The measurements agreed well with those found in a single radial immunodiffusion (SRID) assay. In addition FITC-labeled protein G-tagged anti-h-IgG1 antibodies were used to detect the specific antigen of the involved antibody, namely, h-IgG1, in human sera.
ASJC Scopus subject areas
- Chemistry(all)
- Analytical Chemistry
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In: Analytical chemistry, Vol. 66, No. 22, 15.11.1994, p. 4027-4033.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Fluorescein Isothiocyanate-Labeled Protein G as an Affinity Ligand in Affinity/Immunocapillary Electrophoresis with Fluorescence Detection
AU - Relf, Oscar Werner
AU - Freitag, Ruth
AU - Lausch, Ralf
AU - Scheper, Thomas
PY - 1994/11/15
Y1 - 1994/11/15
N2 - Antibodies from human sera (h-IgG) were tagged with a fluorescent dye (fluorescein isothiocyanate, FITC) through the affinity reaction of FITC-labeled protein G with the Fc fragment of the antibodies. The complexes were quantified by capillary zone electrophoresis (CZE) within 1 min, i.e., fast enough to prevent their dissociation during the measurement. Conditions for the affinity reaction and the CZE analysis could thus be optimized independently. When an FITC-labeled protein G concentration of 10−6 mol/L was used, h-IgG concentrations between 10−6 and 10−9 mol/L were reproducibly quantified (STD < 2%), using an LIF detector. A correlation coefficient, r2, of 0.9988 was established between the peak height and the IgG concentration. Alternatively, h–IgG containing serum samples and the FITC-labeled protein G were simply injected into the CE capillary in consecutive zones, followed by the application of the electrical field. Within 2 min, the affinity complexes were resolved and the IgG content of the serum quantified (r2 = 0.9986). The injection sequence was of no consequence. The measurements agreed well with those found in a single radial immunodiffusion (SRID) assay. In addition FITC-labeled protein G-tagged anti-h-IgG1 antibodies were used to detect the specific antigen of the involved antibody, namely, h-IgG1, in human sera.
AB - Antibodies from human sera (h-IgG) were tagged with a fluorescent dye (fluorescein isothiocyanate, FITC) through the affinity reaction of FITC-labeled protein G with the Fc fragment of the antibodies. The complexes were quantified by capillary zone electrophoresis (CZE) within 1 min, i.e., fast enough to prevent their dissociation during the measurement. Conditions for the affinity reaction and the CZE analysis could thus be optimized independently. When an FITC-labeled protein G concentration of 10−6 mol/L was used, h-IgG concentrations between 10−6 and 10−9 mol/L were reproducibly quantified (STD < 2%), using an LIF detector. A correlation coefficient, r2, of 0.9988 was established between the peak height and the IgG concentration. Alternatively, h–IgG containing serum samples and the FITC-labeled protein G were simply injected into the CE capillary in consecutive zones, followed by the application of the electrical field. Within 2 min, the affinity complexes were resolved and the IgG content of the serum quantified (r2 = 0.9986). The injection sequence was of no consequence. The measurements agreed well with those found in a single radial immunodiffusion (SRID) assay. In addition FITC-labeled protein G-tagged anti-h-IgG1 antibodies were used to detect the specific antigen of the involved antibody, namely, h-IgG1, in human sera.
UR - http://www.scopus.com/inward/record.url?scp=0028774682&partnerID=8YFLogxK
U2 - 10.1021/ac00094a027
DO - 10.1021/ac00094a027
M3 - Article
C2 - 7810902
AN - SCOPUS:0028774682
VL - 66
SP - 4027
EP - 4033
JO - Analytical chemistry
JF - Analytical chemistry
SN - 0003-2700
IS - 22
ER -