A competitive ELISA for the quantitative determination of the novel anti-HIV drug candidate CIGB-210 in biological fluids

• Published in: Journal of immunoassay & immunochemistry 2019-03, Vol.40 (2), p.193-213
• Main Authors: Duarte, Carlos A., Chávez, Sheila, Masforrol, Yordanka, Puertas, Samy, Paneque, Taimí, Ramirez, Anna C., Casillas, Dionne, Puente, Pedro, Garay, Hilda, Fernández-Ortega, Celia
• Format: Article
• Language: English
• Subjects: Analytical methods; Animals; Anti-HIV Agents - analysis; Anti-HIV Agents - blood; Anti-HIV Agents - chemical synthesis; Anti-HIV Agents - chemistry; Assaying; Blood plasma; Body Fluids - chemistry; CIGB-210; Coefficient of variation; competitive ELISA; Enzyme-Linked Immunosorbent Assay; Feasibility studies; Filaments; Freeze thaw cycles; Freeze-thawing; HIV; Human immunodeficiency virus; Humans; Intermediate filaments; Mathematical analysis; Metabolites; Mice; Optimization; Peptides - analysis; Peptides - blood; Peptides - chemical synthesis; Peptides - chemistry; Pharmacology; plasma; Sensitivity analysis; variability; Vimentin
• ISSN: 1532-1819; 1532-4230
• DOI: 10.1080/15321819.2018.1547975

The synthetic peptide CIGB-210 is a promising anti-HIV drug candidate shown to inhibit HIV replication in MT4 cells at the nanomolar range by triggering the rearrangement of vimentin intermediate filaments. Sensitive and specific analytical methods are required for pharmacological studies of CIBG-210 in animals. In this study, we describe the development of a competitive ELISA for the quantitative determination of CIGB-210 using an anti-CIGB-210 hyperimmune serum. After optimization of all the steps, the assay exhibited a dynamic range from 11.87 to 0.0095 µg/mL. The intra-assay coefficient of variation (CV) was lower than or close to 5% for all the six concentrations of the calibrator, and the inter-assay CV was below 10% in five out of the six concentrations tested. No interference of either murine or human plasma was observed. The analyte was stable in plasma after five freeze-thaw cycles, while the hyperimmune serum maintained its binding capacity after 10 freeze-thaw cycles. Furthermore, the ELISA was able to detect the two main metabolites of CIGB-210, although with a tenfold decrease in sensitivity. Our results demonstrate the utility and feasibility of this analytical method for pharmacological experiments in animals as humans.