Abstract
A new solid-phase immunoassay technique has been applied for anti-digoxin monoclonal antibodies (Mabs) detection. At the first assay step, anti-digoxin Mabs (IgG1, Kaff = 9.2 × 109 M-1) were bound to a specially prepared immunosorbent, the microtiter plates coated with digoxin-human serum albumin conjugate (Dig-HSA), in which free amino groups were protected by a glutaraldehyde cross-linking modification. The modification did not essentially influence the antibody-binding capacity of the immunosorbent. After antigen-antibody reaction, free amino groups were located only on the anti-digoxin Mabs, bound to chemically modified immunosorbent. At the second assay step, free amino groups of anti-digoxin Mabs were biotinylated by N-hydroxysuccinimide-biotin ester. Then the biotin residues were detected by the streptavidin-peroxidase conjugate. The method does not require second labeled antibodies and may be used for anti-hapten hybridoma screening.
| Original language | English |
|---|---|
| Pages (from-to) | 235-239 |
| Number of pages | 5 |
| Journal | Immunology Letters |
| Volume | 41 |
| Issue number | 2-3 |
| DOIs | |
| Publication status | Published - 1994 |
| Externally published | Yes |
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Keywords
- Digoxin
- Group-selective immunoassay
- Monoclonal antibody
- Streptavidin-biotin system
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology
Cite this
Group-selective immunoassay. / Yazynin, Sergey A.; Deyev, Sergey M.
In: Immunology Letters, Vol. 41, No. 2-3, 1994, p. 235-239.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Group-selective immunoassay
AU - Yazynin, Sergey A.
AU - Deyev, Sergey M.
PY - 1994
Y1 - 1994
N2 - A new solid-phase immunoassay technique has been applied for anti-digoxin monoclonal antibodies (Mabs) detection. At the first assay step, anti-digoxin Mabs (IgG1, Kaff = 9.2 × 109 M-1) were bound to a specially prepared immunosorbent, the microtiter plates coated with digoxin-human serum albumin conjugate (Dig-HSA), in which free amino groups were protected by a glutaraldehyde cross-linking modification. The modification did not essentially influence the antibody-binding capacity of the immunosorbent. After antigen-antibody reaction, free amino groups were located only on the anti-digoxin Mabs, bound to chemically modified immunosorbent. At the second assay step, free amino groups of anti-digoxin Mabs were biotinylated by N-hydroxysuccinimide-biotin ester. Then the biotin residues were detected by the streptavidin-peroxidase conjugate. The method does not require second labeled antibodies and may be used for anti-hapten hybridoma screening.
AB - A new solid-phase immunoassay technique has been applied for anti-digoxin monoclonal antibodies (Mabs) detection. At the first assay step, anti-digoxin Mabs (IgG1, Kaff = 9.2 × 109 M-1) were bound to a specially prepared immunosorbent, the microtiter plates coated with digoxin-human serum albumin conjugate (Dig-HSA), in which free amino groups were protected by a glutaraldehyde cross-linking modification. The modification did not essentially influence the antibody-binding capacity of the immunosorbent. After antigen-antibody reaction, free amino groups were located only on the anti-digoxin Mabs, bound to chemically modified immunosorbent. At the second assay step, free amino groups of anti-digoxin Mabs were biotinylated by N-hydroxysuccinimide-biotin ester. Then the biotin residues were detected by the streptavidin-peroxidase conjugate. The method does not require second labeled antibodies and may be used for anti-hapten hybridoma screening.
KW - Digoxin
KW - Group-selective immunoassay
KW - Monoclonal antibody
KW - Streptavidin-biotin system
UR - http://www.scopus.com/inward/record.url?scp=0028060588&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028060588&partnerID=8YFLogxK
U2 - 10.1016/0165-2478(94)90139-2
DO - 10.1016/0165-2478(94)90139-2
M3 - Article
VL - 41
SP - 235
EP - 239
JO - Immunology Letters
JF - Immunology Letters
SN - 0165-2478
IS - 2-3
ER -