title: Evaluation of chemically modified carrier proteins for developing monoclonal antibodies against a clinically relevant mutation of cKIT.
abstract: In this report we show that combining double-chemically modified carrier proteins and hetero-functional cross-linkers allows preparing tailor-made hapten-protein carrier conjugates. Accordingly, a new carrier protein has been designed where carboxylic groups were transformed into highly reactive primary amino groups by reaction with ethylendiamine after activation with EDCI. The aminated protein carrier is then modified by different cross-linkers (hyper-activated proteins) at different conditions in order to control the conjugation ratio from 1 to >12 molecules of hapten per carrier protein. Finally, this novel strategy has been successfully used to develop antibodies against a short specific peptide corresponding to a point mutation (D816V) of cKIT, which is a clinically relevant mutation related to mastocytosis and gastrointestinal stroma tumor.
authors: ImmunoStep SL, Salamanca, Spain.
authors: Ricardo
authors: R
authors: Jara-Acevedo
authors: Maria
authors: M
authors: Gonzalez-Gonzalez
authors: María
authors: M
authors: Jara-Acevedo
authors: Jose
authors: J
authors: Claros
authors: Alfredo
authors: A
authors: Conde
authors: Ricardo
authors: R
authors: López-Perez
authors: Alberto
authors: A
authors: Orfao
authors: Manuel
authors: M
authors: Fuentes
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002352:Carrier Proteins / Q000737:chemistry / Q000276:immunology*; D002417:Cattle; D003432:Cross-Linking Reagents / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005029:Ethylenediamines / Q000737:chemistry; D046152:Gastrointestinal Stromal Tumors / Q000235:genetics / Q000276:immunology; D018796:Immunoconjugates / Q000737:chemistry / Q000276:immunology; D008301:Maleimides / Q000737:chemistry; D008415:Mastocytosis / Q000235:genetics / Q000276:immunology; D051379:Mice; D008956:Models, Chemical; D008958:Models, Molecular; D015394:Molecular Structure; D020125:Mutation, Missense; D010455:Peptides / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation; D019009:Proto-Oncogene Proteins c-kit / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D003432:Cross-Linking Reagents; D005029:Ethylenediamines; D018796:Immunoconjugates; D008301:Maleimides; D010455:Peptides; C043592:maleimide; D012710:Serum Albumin, Bovine; D019009:Proto-Oncogene Proteins c-kit