title: Development, validation and use of ELISA for antibodies to human alpha-1 antitrypsin.
abstract: Evaluation of human antibody responses to alpha-1 antitrypsin (AAT) in clinical trials and clinical practice has been limited by the lack of a validated assay. Here we describe the development and validation of an ELISA method for quantification of human and nonhuman primate antibody responses to human AAT. A reference anti-human AAT serum standard was generated using sera from a cynomolgus macaque injected with a recombinant adeno-associated virus vector expressing human AAT. The ELISA was validated for use with human serum dilutions as low as 1:10 and was able to distinguish between specific responses in cynomolgus serum and non-specific increases in apparent antibody titer in serum from subjects in a clinical trial of an AAT gene therapy vector.
authors.affiliation: Applied Genetic Technologies Corp., Alachua, FL 32615, USA.
authors.forename: Guo-Jie
authors.initials: GJ
authors.lastname: Ye
authors.forename: Regina A
authors.initials: RA
authors.lastname: Oshins
authors.forename: Farshid N
authors.initials: FN
authors.lastname: Rouhani
authors.forename: Mark L
authors.initials: ML
authors.lastname: Brantly
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Chulay
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D006801:Humans; D008252:Macaca fascicularis; D000515:alpha 1-Antitrypsin / Q000276:immunology*; D019896:alpha 1-Antitrypsin Deficiency / Q000276:immunology*
chemical_list: D000906:Antibodies; D000515:alpha 1-Antitrypsin