1. Plaque antibody selection: rapid immunological analysis of a large number of recombinant phage clones positive to sera raised against Plasmodium falciparum antigens
- Author
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Luiz Pereira da Silva, Luiz Shozo Ozaki, Thierry Blisnick, Denise Mattei, Micheline Guillotte, Odile Puijalon, Pierre Druihle, Moncef Jendoubi, Institut Pasteur [Paris], Institut Pasteur [Paris] (IP), CHU Pitié-Salpêtrière [AP-HP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)
- Subjects
MESH: Rabbits ,law.invention ,0302 clinical medicine ,law ,MESH: Immune Sera ,Immunology and Allergy ,MESH: Animals ,Cloning, Molecular ,MESH: Plasmodium falciparum ,ComputingMilieux_MISCELLANEOUS ,0303 health sciences ,medicine.diagnostic_test ,Collodion ,3. Good health ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,MESH: DNA, Recombinant ,Recombinant DNA ,Electrophoresis, Polyacrylamide Gel ,MESH: Paper ,Rabbits ,Antibody ,Clone (B-cell biology) ,Adult ,Paper ,MESH: Collodion ,Plasmodium falciparum ,030231 tropical medicine ,Immunology ,DNA, Recombinant ,Antigens, Protozoan ,Biology ,Molecular cloning ,03 medical and health sciences ,Antigen ,Western blot ,parasitic diseases ,medicine ,Animals ,Humans ,MESH: Cloning, Molecular ,MESH: T-Phages ,030304 developmental biology ,Antiserum ,MESH: Humans ,Immune Sera ,Immunoselection ,MESH: Adult ,Expression library ,biology.organism_classification ,DNA library ,Virology ,biology.protein ,T-Phages ,MESH: Antigens, Protozoan ,MESH: Electrophoresis, Polyacrylamide Gel - Abstract
International audience; A library of Plasmodium falciparum genomic DNA on the lambda gt11 phage vector was screened for clones positive to a rabbit serum raised against a purified fraction of P. falciparum proteins and a pool of sera from malaria patients. The positive clones were characterized with antibodies purified by the plaque antibody selection technique. This technique consist of purifying specific antibodies on a nitrocellulose filter blotted directly on a lawn of plaques of an antigen-producing phage clone. The purified antibodies are then used as a probe in a Western blot of parasite protein extract, for preliminary characterization of the clones. Using this method, two different clones coding for P. falciparum antigens were identified with the rabbit serum and about 20 with the human sera. This method can be of general use, i.e. it is not limited to parasite systems, and facilitates the immunological analysis and identification of a large number of clones.
- Published
- 1986