title: Production, safety and antitumor efficacy of recombinant Oncofetal Antigen/immature laminin receptor protein.
abstract: We describe here for the first time an efficient high yield production method for clinical grade recombinant human Oncofetal Antigen/immature laminin receptor protein (OFA/iLRP). We also demonstrate significant antitumor activity for this protein when administered in liposomal delivery form in a murine model of syngeneic fibrosarcoma. OFA/iLRP is a therapeutically very promising universal tumor antigen that is expressed in all mammalian solid tumors tested so far. We have cloned the human OFA/iLRP cDNA in a bacterial expression plasmid which incorporates a 6x HIS-tag. Large scale cultures of the plasmid transformed Escherichia coli were performed and the crude HIS-tagged OFA/iLRP was isolated as inclusion bodies and solubilized in guanidine chloride. The protein was then purified by successive passage through three column chromatography steps of immobilized metal affinity, anion exchange, and gel filtration. The resulting protein was 94% pure and practically devoid of endotoxin and host cell protein. The purified OFA/iLRP was tested in mice for safety and efficacy in tumor rejection with satisfactory results. This protein will be used for loading onto autologous dendritic cells in an FDA approved phase I/II human cancer vaccine trial in OFA/iLRP-positive breast cancer patients.
authors.affiliation: Department of Microbiology and Immunology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA. abarsoum@jaguar1.usouthal.edu
authors.forename: Adel L
authors.initials: AL
authors.lastname: Barsoum
authors.forename: Bainan
authors.initials: B
authors.lastname: Liu
authors.forename: James W
authors.initials: JW
authors.lastname: Rohrer
authors.forename: Joseph H
authors.initials: JH
authors.lastname: Coggin
authors.forename: J Allan
authors.initials: JA
authors.lastname: Tucker
authors.forename: Lewis K
authors.initials: LK
authors.lastname: Pannell
authors.forename: Paul O
authors.initials: PO
authors.lastname: Schwarzenberger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D019496:Cancer Vaccines / Q000009:adverse effects / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor / Q000276:immunology; D002986:Clinical Trials as Topic; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D005354:Fibrosarcoma / Q000276:immunology* / Q000378:metabolism; D019791:Guanidine / Q000494:pharmacology; D006639:Histidine / Q000737:chemistry; D006801:Humans; D002479:Inclusion Bodies / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids / Q000235:genetics; D011786:Quality Control; D017474:Receptors, Laminin / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D012995:Solubility; D014169:Transformation, Bacterial; D023041:Xenograft Model Antitumor Assays
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; C510624:RPSA protein, human; D017474:Receptors, Laminin; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C031192:oncofetal antigens; D006639:Histidine; D019791:Guanidine