Concentration of progesterone monoclonal antibodies using different non-chromatographic methods

Abstract

Hybridoma supernatants containing MAbs against progesterone (P₄) were concentrated using sephadex 0-10, dialysorb, polyethylene glycol (PEG) 20000, Centriprep, PEG 6000 and saturated ammonium sulphate. Total protein concentration was estimated by Lowry's method. Specific protein yield was estimated using capture ELISA by diluting the concentrates to pre-concentration level. The mean specific protein yields were 85,65,70,71 and 59% with sephadex 0-10, centriprep, dialysorb, saturated ammonium sulphate and PEG 6000 respectively. No recovery of specific protein was obtained with PEG 20000. Based on the results, concentration of P₄ MAbs using sephadex 0-10 appears to be an ideal, simple and rapid procedure for maximum recovery of P₄ specific MAbs.