Exploiting light chains for the scalable generation and platform purification of native human bispecific IgG.

Bispecific antibodies enable unique therapeutic approaches but it remains a challenge to produce them at the industrial scale, and the modifications introduced to achieve bispecificity often have an impact on stability and risk of immunogenicity. Here we describe a fully human bispecific IgG devoid of any modification, which can be produced at the industrial scale, using a platform process. This format, referred to as a κλ-body, is assembled by co-expressing one heavy chain and two different light chains, one κ and one λ.

Size exclusion-high-performance liquid chromatography (SEC-HPLC).

The size exclusion-high-performance liquid chromatography is a high-throughput analytical method, through isocratic condition, that allows to determine and quantify the level of aggregates and fragments of purified antibodies. Here, we describe the preparation of the samples, chromatographic conditions to be used (buffer preparation, quantity injected, flow rate, run time, column suitability, etc.), validity of the analysis, and integration of the chromatogram in order to calculate the proportion of the different components.

Ion exchange-high-performance liquid chromatography (IEX-HPLC).

The ion exchange-high-performance liquid chromatography is a high-throughput analytical method that allows to determine the charge profile of purified antibodies. Here, we describe the preparation of the samples, chromatographic conditions to be used (buffer preparation, salt gradient, quantity injected, flow rate, run time, column suitability, etc.), validity of the analysis, and integration of the chromatogram in order to calculate the proportion of the different isoforms.