An artificial protein L for the purification of immunoglobulins and fab fragments by affinity chromatography.
The development and characterization of an artificial protein L (PpL) for the affinity purification of antibodies is described. Ligand 8/7, which emerged as the lead from a de novo designed combinatorial library of ligands, inhibits the interaction of PpL with IgG and Fab by competitive ELISA and shows negligible binding to Fc. The ligand 8/7 adsorbent (Ka approximately 10(4) M(-1)) compared well with PpL in binding to immunoglobulins from different classes and sources and, in addition, bound to IgG1 with K and lambda isotypes (92% and 100% of loaded protein) and polyclonal IgG from sheep, cow, goat and chicken. These properties were also reflected in the efficient isolation of immunoglobulins from crude samples.
Roque
A Cecília A
AC
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, 1049-001 Lisboa, Portugal.
Taipa
M Angela
MA
Lowe
Christopher R
CR
D001426:Bacterial Proteins / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D006863:Hydrogen-Ion Concentration; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification*; D007136:Immunoglobulins / Q000302:isolation & purification*; D009994:Osmolar Concentration
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D007140:Immunoglobulin Fab Fragments; D007136:Immunoglobulins; C116593:L-protein, Peptococcus magnus