Three-step purification of bacterially expressed human single-chain Fv antibodies for clinical applications

We have obtained a cell line which secretes a human monoclonal IgM (B7) rea cting with the myosin heavy chain of human heart. We have constructed singl e-chain fragments (scFv) of B7. The scFv may be useful for the imaging of m yocardial necrosis after myocarditis, cardiac drug toxicosis or graft rejec tion. The aim of our work was to purify the scFv for immunoscintigraphy. We describe several purification steps including immobilized metal affinity c hromatography (IMAC), anti-c-myc monoclonal antibody affinity chromatograph y, size-exclusion chromatography with Superdex(R) 75 HR 10/30 and ion-excha nge chromatography (mini Q TM 30Q). (C) 2000 Elsevier Science B.V. All righ ts reserved.