Abteilung Biotechnologie, Institut für Biochemie und Biotechnologie, Technische Universität Braunschweig, Braunschweig, Germany.
MAbs. 2010 Jan-Feb;2(1):73-6. doi: 10.4161/mabs.2.1.10784. Epub 2010 Jan 26.
Assembly of immunoglobulin G (IgG) molecules from two heavy and two light chains can be facilitated by connecting the light chain to the heavy chain by an oligopeptide linker. Production of the anti-lysozyme D1.3-single chain (sc) IgG1 in HEK293T cells yielded up to 8 mg/L functional scIgG polypeptide. Size exclusion chromatography of material purified by protein-A affinity chromatography revealed that the majority of the D1.3-scIgG1 molecules were 150 kDa monomers, with a K(D) of 1.8 x 10(-10) M measured by surface plasmon resonance; however, significant fractions of scIgG dimers and oligomers with molecular masses of 300 kDa and >600 kDa, respectively, were identified. The oligomerization resulted in an increased avidity. The observed oligomerization capability may allow new approaches for the generation of bispecific/multivalent antibodies.
免疫球蛋白 G（IgG）分子由两条重链和两条轻链组成，通过将轻链与重链连接起来可以促进其组装。通过寡肽接头将抗溶菌酶 D1.3-单链（sc）IgG1 连接到 HEK293T 细胞中，可生产出高达 8 mg/L 的功能性 scIgG 多肽。通过蛋白 A 亲和层析纯化的材料的分子筛层析表明，大多数 D1.3-scIgG1 分子是 150 kDa 的单体，通过表面等离子体共振（surface plasmon resonance）测定的 K(D)值为 1.8 x 10(-10) M；然而，鉴定出具有 300 kDa 和 >600 kDa 分子量的 scIgG 二聚体和多聚体的显著分数。二聚化导致亲和力增加。观察到的多聚化能力可能为双特异性/多价抗体的产生提供新的方法。
