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Novel approach to activity evaluation for release-active forms of anti-interferon-gamma antibodies based on enzyme-linked immunoassay.

Gavrilova ES, Bobrovnik SA, Sherriff G, Myslivets AA, Tarasov SA, Epstein OI - PLoS ONE (2014)

Bottom Line: In this study, the enzyme-linked immunosorbent assay was utilized to examine the effect of RA forms of Abs specific for human interferon gamma on the interaction between monoclonal anti-interferon gamma antibodies and recombinant human interferon gamma.These data demonstrated the importance of using relatively low concentrations of both soluble and plate-immobilized interferon gamma to detect the effects of RA forms of Abs to interferon gamma on the binding of monoclonal antibodies to interferon gamma.It has been suggested that the observed influence of RA forms of Abs on 'antibody-antigen' interaction could be used to detect and analyze the activity of drugs containing RA forms of Abs.

View Article: PubMed Central - PubMed

Affiliation: OOO "NPF "MATERIA MEDICA HOLDING", Moscow, Russian Federation.

ABSTRACT
Selection of a suitable assay to measure the activity of drug agents based on release-active forms of anti-interferon-gamma antibodies (RA forms of Abs) is an important step forward in the investigation of such agents. In this study, the enzyme-linked immunosorbent assay was utilized to examine the effect of RA forms of Abs specific for human interferon gamma on the interaction between monoclonal anti-interferon gamma antibodies and recombinant human interferon gamma. The experimental data and the results obtained by using relevant mathematical analysis showed that such RA forms of Abs are able to modulate the monoclonal antibody interaction with both soluble and immobilized (to the assay plate well) interferon gamma. These data demonstrated the importance of using relatively low concentrations of both soluble and plate-immobilized interferon gamma to detect the effects of RA forms of Abs to interferon gamma on the binding of monoclonal antibodies to interferon gamma. It has been suggested that the observed influence of RA forms of Abs on 'antibody-antigen' interaction could be used to detect and analyze the activity of drugs containing RA forms of Abs.

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Linear relationship between mAbs to IFN-gamma dilutions and optical density.The concentration of IFN-gamma absorbed on the plate is 1 µg/ml. Dilutions of mAbs are on X axis (where 0.00001 means dilution of starting antibodies to 1∶100000 times, 0.00002 means dilution of starting antibodies to 1∶50000 times etc.). Optical densities are average values of three measurements. The error bars are the standard deviations.
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pone-0097017-g001: Linear relationship between mAbs to IFN-gamma dilutions and optical density.The concentration of IFN-gamma absorbed on the plate is 1 µg/ml. Dilutions of mAbs are on X axis (where 0.00001 means dilution of starting antibodies to 1∶100000 times, 0.00002 means dilution of starting antibodies to 1∶50000 times etc.). Optical densities are average values of three measurements. The error bars are the standard deviations.

Mentions: The experiment was performed in accordance with the standard ELISA protocol. For this experiment only mAbs without RA forms of Abs to IFN-gamma or control were used as test samples. Dilutions of mAbs were 1∶80000 (0,0000125), 1∶40000 (0,000025) and 1∶20000 (0,00005). The concentration of IFN-gamma adsorbed onto the plate was 1 µg/ml. An example of concentration/color relationship is presented in Fig. 1.


Novel approach to activity evaluation for release-active forms of anti-interferon-gamma antibodies based on enzyme-linked immunoassay.

Gavrilova ES, Bobrovnik SA, Sherriff G, Myslivets AA, Tarasov SA, Epstein OI - PLoS ONE (2014)

Linear relationship between mAbs to IFN-gamma dilutions and optical density.The concentration of IFN-gamma absorbed on the plate is 1 µg/ml. Dilutions of mAbs are on X axis (where 0.00001 means dilution of starting antibodies to 1∶100000 times, 0.00002 means dilution of starting antibodies to 1∶50000 times etc.). Optical densities are average values of three measurements. The error bars are the standard deviations.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4016219&req=5

pone-0097017-g001: Linear relationship between mAbs to IFN-gamma dilutions and optical density.The concentration of IFN-gamma absorbed on the plate is 1 µg/ml. Dilutions of mAbs are on X axis (where 0.00001 means dilution of starting antibodies to 1∶100000 times, 0.00002 means dilution of starting antibodies to 1∶50000 times etc.). Optical densities are average values of three measurements. The error bars are the standard deviations.
Mentions: The experiment was performed in accordance with the standard ELISA protocol. For this experiment only mAbs without RA forms of Abs to IFN-gamma or control were used as test samples. Dilutions of mAbs were 1∶80000 (0,0000125), 1∶40000 (0,000025) and 1∶20000 (0,00005). The concentration of IFN-gamma adsorbed onto the plate was 1 µg/ml. An example of concentration/color relationship is presented in Fig. 1.

Bottom Line: In this study, the enzyme-linked immunosorbent assay was utilized to examine the effect of RA forms of Abs specific for human interferon gamma on the interaction between monoclonal anti-interferon gamma antibodies and recombinant human interferon gamma.These data demonstrated the importance of using relatively low concentrations of both soluble and plate-immobilized interferon gamma to detect the effects of RA forms of Abs to interferon gamma on the binding of monoclonal antibodies to interferon gamma.It has been suggested that the observed influence of RA forms of Abs on 'antibody-antigen' interaction could be used to detect and analyze the activity of drugs containing RA forms of Abs.

View Article: PubMed Central - PubMed

Affiliation: OOO "NPF "MATERIA MEDICA HOLDING", Moscow, Russian Federation.

ABSTRACT
Selection of a suitable assay to measure the activity of drug agents based on release-active forms of anti-interferon-gamma antibodies (RA forms of Abs) is an important step forward in the investigation of such agents. In this study, the enzyme-linked immunosorbent assay was utilized to examine the effect of RA forms of Abs specific for human interferon gamma on the interaction between monoclonal anti-interferon gamma antibodies and recombinant human interferon gamma. The experimental data and the results obtained by using relevant mathematical analysis showed that such RA forms of Abs are able to modulate the monoclonal antibody interaction with both soluble and immobilized (to the assay plate well) interferon gamma. These data demonstrated the importance of using relatively low concentrations of both soluble and plate-immobilized interferon gamma to detect the effects of RA forms of Abs to interferon gamma on the binding of monoclonal antibodies to interferon gamma. It has been suggested that the observed influence of RA forms of Abs on 'antibody-antigen' interaction could be used to detect and analyze the activity of drugs containing RA forms of Abs.

Show MeSH
Related in: MedlinePlus