title: Specific detection and quantitation of bovine IgG in bioreactor derived mouse mAb preparations.
abstract: Monoclonal antibody and recombinant protein production benefits greatly from bovine serum as an additive. The caveat is that bovine serum IgG, co-purifies with mAbs and IgG Fc-containing fusion proteins and it presents a contaminant in the end products. In order to analytically validate the products, species specific reagents are needed that react with bovine IgG exclusively. Our attempts to find such commercially available reagents failed. Here, we report the production of species specific mAbs which recognize bovine IgG even in the presence of excess amount of mouse IgG. We present five mAbs: Bsi4028, Bsi4032, Bsi4033, Bsi4034 and Bsi4035 suitable to determine the presence of bovine IgG contamination via ELISA or immunoblotting in bioreactor derived mouse mAb preparations. To quantitate bovine IgG content we developed sensitive sandwich ELISAs capable to detect bovine IgG contaminant in the ng/ml (~10-11M/l) range. Finally, we show that bovine IgG is efficiently removed from bioreactor produced mouse mAb preparation via affinity depletion columns prepared with Bsi4028, Bsi4032, Bsi4033, Bsi4034, Bsi4035 mAbs.
authors: Biosystems International Kft, 98 Nagyerdei krt 4032, Debrecen, Hungary. Electronic address: anna.debreceni@biosys-intl.com.
authors: Anna
authors: A
authors: Gall-Debreceni
authors: Biosystems International Kft, 98 Nagyerdei krt 4032, Debrecen, Hungary; Laboratory of Monoclonal Antibody Proteomics, Department of Human Genetics, Faculty of Medicine, University of Debrecen, Debrecen, Budapest, Hungary.
authors: Jozsef
authors: J
authors: Lazar
authors: Biosystems International Kft, 98 Nagyerdei krt 4032, Debrecen, Hungary.
authors: Janos
authors: J
authors: Kadas
authors: Biosystems International Kft, 98 Nagyerdei krt 4032, Debrecen, Hungary.
authors: Attila
authors: A
authors: Balogh
authors: Biosystems International Kft, 98 Nagyerdei krt 4032, Debrecen, Hungary.
authors: Annamaria
authors: A
authors: Ferenczi
authors: Budapest Zoo and Botanical Garden, Budapest, Hungary.
authors: Endre
authors: E
authors: Sos
authors: Biosystems International Kft, 98 Nagyerdei krt 4032, Debrecen, Hungary; Laboratory of Monoclonal Antibody Proteomics, Department of Human Genetics, Faculty of Medicine, University of Debrecen, Debrecen, Budapest, Hungary.
authors: Laszlo
authors: L
authors: Takacs
authors: Biosystems International Kft, 98 Nagyerdei krt 4032, Debrecen, Hungary.
authors: Istvan
authors: I
authors: Kurucz
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000032:analysis*; D019149:Bioreactors; D002417:Cattle; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology; D005260:Female; D006825:Hybridomas / Q000276:immunology*; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000032:analysis*; D013045:Species Specificity
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000939:Epitopes; D007074:Immunoglobulin G; D011994:Recombinant Proteins
keywords: Bovine IgG contamination; Detection; Monoclonal antibody; Purification