Expression of a VHCκ chimaeric protein in mouse myeloma cells

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Abstract
The heavy (H) and light (L) chains of antibodies consist of variable (V) and constant (C) regions. The V regions of the heavy and light chains form the antibody combining site1,2. To determine whether a V region could be functional when joined to a polypeptide other than its own C region, we constructed a chimaeric gene encoding the V region of a mouse heavy chain and the C region of a mouse κ light chain (VHCκ). The heavy-chain gene is derived from an A/J mouse hybridoma cell line 36-65 whose antibody product (γ1, κ) is specific for the hapten azophenylarsonate3. We report here that, when introduced into a mouse myeloma cell line, the chimaeric gene is expressed and a protein of the expected molecular weight is secreted into the medium. As light chains tend to dimerize4,5 we expected that the VHCκ protein might associate with light chain from the cell line 36-65 to form an antibody-binding molecule. Affinity binding experiments and Ka determination indicate that this is the case. Dimers of this type offer a novel and interesting alternative to existing antibody-binding molecules.