Oligomeric forms of single chain immunoglobulin (scIgG).
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.
Schirrmann
Thomas
T
Abteilung Biotechnologie, Institut für Biochemie und Biotechnologie, Technische Universität Braunschweig, Braunschweig, Germany. th.schirrmann@tu-bs.de
Menzel
Christian
C
Hust
Michael
M
Prilop
Jessica
J
Jostock
Thomas
T
Dübel
Stefan
S
D000818:Animals; D000915:Antibody Affinity; D002460:Cell Line; D002846:Chromatography, Affinity; D006801:Humans; D051379:Mice; D009113:Muramidase / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D015202:Protein Engineering; D055503:Protein Multimerization; D017452:Receptors, IgG / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D020349:Surface Plasmon Resonance
C489017:FCGR1A protein, human; D010446:Peptide Fragments; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D009113:Muramidase