Details
| Original language | English |
|---|---|
| Pages (from-to) | 636-641 |
| Number of pages | 6 |
| Journal | ELECTROPHORESIS |
| Volume | 16 |
| Issue number | 1 |
| Publication status | Published - 1995 |
| Externally published | Yes |
Abstract
A method for the rapid and sensitive determination of immunoglobulin G (IgG) in cultivation media by an affinity assay using capillary electrophoresis is presented. For that purpose we evaluated protein A conjugated with a fluorescent dye such as fluorescein diisocyanate or dichlorotriazinyl‐aminofluorescein as an affinity ligand. The ligand formed a fluorescing complex with immunoglobulin G in the sample and rapid separation from excess protein A was performed by capillary zone electrophoresis. However, only partial resolution of the zones was achieved when protein A as a whole molecule was utilized. In contrast, baseline resolution of the zones was obtained when recombinant fragments of protein A were used as affinity ligands. Immunoglobulin concentrations in the range of two orders of magnitude were determined. Due to the specifity of protein A for immunoglobulin G, analysis can be carried out even in the presence of high concentrations of other components and in cultivation media. Thus, the capillary electrophoretic affinity assay was successfully applied to monitor monoclonal antibodies in a cultivation process.
Keywords
- Capillary electrophoretic affinity assay, Immunoglobulin G, Laser‐induced fluorescence detection, Protein A, Recombinant proteins
ASJC Scopus subject areas
- Chemistry(all)
- Analytical Chemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Clinical Biochemistry
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In: ELECTROPHORESIS, Vol. 16, No. 1, 1995, p. 636-641.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Analysis of immunoglobulin G using a capillary electrophoretic affinity assay with protein A and laser‐induced fluorescence detection
AU - Lausch, Ralf
AU - Reif, Oscar‐Werner ‐W
AU - Riechel, Peter
AU - Scheper, Thomas
PY - 1995
Y1 - 1995
N2 - A method for the rapid and sensitive determination of immunoglobulin G (IgG) in cultivation media by an affinity assay using capillary electrophoresis is presented. For that purpose we evaluated protein A conjugated with a fluorescent dye such as fluorescein diisocyanate or dichlorotriazinyl‐aminofluorescein as an affinity ligand. The ligand formed a fluorescing complex with immunoglobulin G in the sample and rapid separation from excess protein A was performed by capillary zone electrophoresis. However, only partial resolution of the zones was achieved when protein A as a whole molecule was utilized. In contrast, baseline resolution of the zones was obtained when recombinant fragments of protein A were used as affinity ligands. Immunoglobulin concentrations in the range of two orders of magnitude were determined. Due to the specifity of protein A for immunoglobulin G, analysis can be carried out even in the presence of high concentrations of other components and in cultivation media. Thus, the capillary electrophoretic affinity assay was successfully applied to monitor monoclonal antibodies in a cultivation process.
AB - A method for the rapid and sensitive determination of immunoglobulin G (IgG) in cultivation media by an affinity assay using capillary electrophoresis is presented. For that purpose we evaluated protein A conjugated with a fluorescent dye such as fluorescein diisocyanate or dichlorotriazinyl‐aminofluorescein as an affinity ligand. The ligand formed a fluorescing complex with immunoglobulin G in the sample and rapid separation from excess protein A was performed by capillary zone electrophoresis. However, only partial resolution of the zones was achieved when protein A as a whole molecule was utilized. In contrast, baseline resolution of the zones was obtained when recombinant fragments of protein A were used as affinity ligands. Immunoglobulin concentrations in the range of two orders of magnitude were determined. Due to the specifity of protein A for immunoglobulin G, analysis can be carried out even in the presence of high concentrations of other components and in cultivation media. Thus, the capillary electrophoretic affinity assay was successfully applied to monitor monoclonal antibodies in a cultivation process.
KW - Capillary electrophoretic affinity assay
KW - Immunoglobulin G
KW - Laser‐induced fluorescence detection
KW - Protein A
KW - Recombinant proteins
UR - http://www.scopus.com/inward/record.url?scp=0028947314&partnerID=8YFLogxK
U2 - 10.1002/elps.11501601102
DO - 10.1002/elps.11501601102
M3 - Article
C2 - 7588537
AN - SCOPUS:0028947314
VL - 16
SP - 636
EP - 641
JO - ELECTROPHORESIS
JF - ELECTROPHORESIS
SN - 0173-0835
IS - 1
ER -