title: Determination of dissociation constants of anti-ALP monoclonal antibodies by an ELISA - based method.
abstract: BACKGROUND
  Dissociation constant (Kd) is of major significance in immunoassay. Since affinity may be influenced by the immunoassay methodology it is important to determine this parameter under the condition of the assay used.
  
  
  OBJECTIVE
  To employ a rapid and simple ELISA- based method for measuring dissociation constants of two Alkaline Phosphatase (ALP) specific MAbs (A1G8F7 and A1G9G3) established in our laboratory.
  
  
  METHODS
  A simple and rapid method on enzyme- linked immunosorbent assay (ELISA) was developed for measuring the dissociation constant of antibody – antigen reactions. In this method the ability of increasing amounts of antigen to bind to antibody measured in the fluid phase.
  
  
  RESULTS
  Based on the data obtained from this study, the dissociation constants of A1G8F7 and A1G9G3 MAbs were 3.8 × 10−9 and 4.3 × 10−9 M, respectively.
  
  
  CONCLUSIONS
  A1G8F7 and A1G9G3 MAbs with reasonably high affinity are suggested for used in very immunoassay such as ELISA, immunocytochemistry and immunihistochemistry (APAAP method).
authors: Department of Immunology, Paramedical School, Birjand University of Medical Sciences, Birjand, Iran.
authors: Mohsen
authors: M
authors: Naseri
authors: Department of Immunology, School of Medicine, Tarbiat Modares University, Birjand, Iran.
authors: Mohammad
authors: M
authors: Moazzeni
authors: Department of Immunology, School of Medicine, Tarbiat Modares University, Birjand, Iran.
authors: Ali Akbar
authors: AA
authors: Purfathollah
authors: Department of Medical Biotechnology School of Medical Sciences, Tarbiat Modares University, Birjand, Iran.
authors: Rahbarizadeh
authors: R
authors: Fatemeh
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology; D000915:Antibody Affinity; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D000941:Antigens / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007422:Intestines / Q000737:chemistry / Q000201:enzymology; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000941:Antigens; D000469:Alkaline Phosphatase
keywords: Dissociation constant; ELISA; alkaline phosphatase; antibody affinity; monoclonal antibody