Chromatographic isolation of cells and other complex biological materials

A chromatography column comprising an affinity chromatography stationary matrix suitable for a CD4+ lymphocyte cell separation,

• wherein the affinity chromatography stationary matrix comprises immobilized thereon both (1) an affinity reagent that comprises a streptavidin mutein and (2) a receptor binding reagent,
• wherein the streptavidin mutein is immobilized on the chromatography stationary matrix,
• wherein the receptor binding reagent comprises:
• (1) a Fab monomer fragment that binds CD4 on the surface of lymphocytes with (i) a dissociation constant (KD) of 10−3 to 10−7 M and/or (b) a dissociation rate constant (koff) of 3×10−5 sec−1 or greater, and (2) a binding partner C comprising a streptavidin-binding peptide fused to the C-terminus of the Fab monomer fragment,
• wherein the binding partner C of the receptor binding reagent is bound to a binding site Z of the immobilized streptavidin mutein of the affinity reagent, thereby immobilizing the receptor binding reagent on the chromatography matrix;
• wherein the Fab fragment comprises a heavy chain comprising the sequence set forth in SEQ ID NO:
• 1 and a light chain comprising the sequence set forth in SEQ ID NO:
• 2,
• wherein the streptavidin-binding peptide comprises the amino acid sequence of SEQ ID NO:
• 13 or SEQ ID NO:
• 3, and
• wherein the binding site Z has a lower dissociation constant (Kd) for biotin or a biotin analog than for the binding partner C.