Evaluation of dissociation constants of antigen-antibody complexes by ELISA

Kim, B.B.; Dikova, E.; Sheller, U.; Dikov, Mikhail M.; Gavrilova, E. M.; Egorov, A. M.

doi:10.1016/0022-1759(90)90192-x

Cited by 23 publications

(8 citation statements)

References 4 publications

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“…Immunoglobulin G from non-immune rabbits was used as a control. Dissociation constants for antibodies were estimated as described (21). ATP hydrolysis was measured as the increase in inorganic phosphate determined colorimetrically at 350 nm in a modified assay of Lin and Morales (22).…”

Section: Methodsmentioning

confidence: 99%

Peptidoglycan Recognition Protein Tag7 Forms a Cytotoxic Complex with Heat Shock Protein 70 in Solution and in Lymphocytes

Sashchenko

Dukhanina

Yashin

et al. 2004

Journal of Biological Chemistry

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The peptidoglycan recognition protein Tag7 is shown to form a stable 1:1 complex with the major stress protein Hsp70. Neither protein is cytotoxic by itself, but their complex induces apoptotic death in several tumorderived cell lines even at subnanomolar concentrations. The minimal part of Hsp70 needed to evoke cytotoxicity is residues 450 -463 of its peptide-binding domain, but full cytotoxicity requires its ATPase activity; remarkably, Tag7 liberated from the complex at high ATP is not cytotoxic. The Tag7-Hsp70 complex is produced by tag7-transfected cells and by lymphokine-activated killers, being assembled within the cell and released into the medium through the Golgi apparatus by a mechanism different from the commonly known granule exocytosis. Thus, we demonstrate how a heat shock protein may perform functions clearly distinct from chaperoning or cell rescue and how peptidoglycan recognition proteins may be involved in innate immunity and anti-cancer defense.

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Section: Methodsmentioning

confidence: 99%

Peptidoglycan Recognition Protein Tag7 Forms a Cytotoxic Complex with Heat Shock Protein 70 in Solution and in Lymphocytes

Sashchenko

Dukhanina

Yashin

et al. 2004

Journal of Biological Chemistry

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“…However, the linearity of Figure 2A signifies study of the biphasic interaction over an antibody concentration range where 2K Ag Ã Ab C Ab ( 1. Such elimination of the binding constant for the biphasic interaction has also been noted by Kim et al (1990).…”

Section: The Original Characterization Of An Antigen-antibody Reactiomentioning

confidence: 83%

Allowance for antibody bivalence in the characterization of interactions by ELISA

Winzor¹

2010

J of Molecular Recognition

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The objective of this review is to remove empiricism from the characterization of immunospecific interactions by enzyme-linked immunosorbent assay (ELISA). In place of the original presumption that the absorbance generated by the enzyme-linked assay could be regarded as a measure of free antibody concentration, the stance is taken that the parameter being monitored is the concentration of antibody complexed with immobilized antigen on the microtiter plate. After the presentation of general binding theory that takes ligand multivalence into account, that theory is adapted to incorporate the simplifying circumstances that prevail in an ELISA study. Validity of the original expressions for characterizing antigen-antibody interactions by competitive ELISA is confirmed, thereby refuting reported concerns about the need for amendment of the theoretical expressions to take into account bivalence of the antibody.

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“…Evaluation of McAbs affinity. The McAb affinity constants were evaluated according to the inhibition method proposed by B. Friguet [12], modified by B. Kim [13]. Affinity constants were calculated by S. Bobrovnik recommendation [14].…”

Section: Methodsmentioning

confidence: 99%

New monoclonal antibodies to the Chlamydia trachomatis main outer membrane protein and their immunobiological properties

Galkin¹,

Besarab²,

Gorshunov³

et al. 2019

Ukr.Biochem.J

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Evaluation of dissociation constants of antigen-antibody complexes by ELISA

Cited by 23 publications

References 4 publications

Peptidoglycan Recognition Protein Tag7 Forms a Cytotoxic Complex with Heat Shock Protein 70 in Solution and in Lymphocytes

Peptidoglycan Recognition Protein Tag7 Forms a Cytotoxic Complex with Heat Shock Protein 70 in Solution and in Lymphocytes

Allowance for antibody bivalence in the characterization of interactions by ELISA

New monoclonal antibodies to the Chlamydia trachomatis main outer membrane protein and their immunobiological properties

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