Preparation of clinical grade monoclonal antibodies from serum-containing cell culture supernatants.

Three mouse monoclonal antibodies (Mab), RIV6, MN12, and WT31, were purified from cell culture supernatants containing foetal bovine serum (FBS) by two-step purification protocols, involving protein A affinity and ion exchange chromatography. Provided that the purification conditions were adapted to the physico-chemical properties of the individual Mab, clinical grade products could be obtained. The residual levels of bovine IgG originating from FBS were below 1% on a protein basis.

A purification strategy for the production of clinical grade monoclonal antibodies.

A strategy for the purification of murine IgG monoclonal antibodies (MAbs) is described. It consists of a combination of protein A affinity chromatography and ion exchange chromatography. The method was used for the purification of two MAbs, WT31 and MN12.

Two-step purification of a murine monoclonal antibody intended for therapeutic application in man. Optimisation of purification conditions and scaling up.

The murine hybridoma cell line WT31, which produces a monoclonal antibody (Mab) of the IgG1 isotype with specificity for the human T cell receptor, was grown in batch-suspension cultures in the presence of foetal bovine serum (FBS). To acquire a clinical grade product for the reversal of allograft rejection, the clarified and concentrated cell culture supernatant was purified by a two-step chromatographic procedure, involving protein A affinity chromatography and Q Sepharose anion exchange chromatography. After choosing the appropriate conditions on a small scale, the purification process was scaled up.