title: Glycomapping the fine specificity of monoclonal and polyclonal Lewis antibodies with type-specific Lewis kodecytes and function-spacer-lipid constructs printed on paper.
abstract: BACKGROUND
  Lewis serologic reagents frequently give inaccurate phenotyping results. Furthermore these serologic reagents are often used in nonserologic assays such as inhibition and immunohistochemistry. In both scenarios knowledge of the fine specificity and cross-reactivity of these reagents will improve the quality of results obtained.
  
  
  METHODS
  A range of contemporary and historical workshop and developmental Lewis reagents including mouse monoclonal (MoAb) and human and goat polyclonal (PoAb) reagents were evaluated. All were evaluated both against Lewis kodecytes expressing only single Le(a) , Le(b) , ALe(b) , BLe(b) , Le(x) , Le(y) , ALe(y) , or BLe(y) antigens and against the same antigens inkjet printed on a paper-based microplate and analyzed by enzyme immunoassay. Nine clinical samples were also evaluated. A kodecyte antigen dilution sensitivity assay was used to establish the ratio of Le(b) antigen between group A1 /A2 and O RBCs.
  
  
  RESULTS
  A continuum of cross-reactivity from Le(x) through to H was observed with MoAbs. All PoAb and few MoAb anti-Le(a) samples and reagents cross-reacted to some degree with Le(b) antigen. Some PoAb and MoAb anti-Le(b) did not cross-react with Le(a) . All polyclonal goat anti-Le(b) reagents showed substantial activity against ALe(b) and BLe(b) , while no MoAb reagent had this activity. A1 RBCs had less than half the Le(b) antigen of A2 /O RBCs.
  
  
  CONCLUSIONS
  Substantial cross-reactivity of both MoAbs and PoAbs with related antigens highlights the risks of using serologic reagents in nonserologic assays or against synthetic antigens. The lack of ALe(b) activity in anti-Le(b) MoAbs explains their poor performance against blood group A1 Le(a-b+) phenotypes.
authors.affiliation: Biotech Innovation Centre, Faculty of Design and Creative Technologies, AUT University, Auckland, New Zealand.
authors.forename: Eleanor
authors.initials: E
authors.lastname: Williams
authors.affiliation: Shemyakin Institute of Bioorganic Chemistry, Moscow, Russian Federation.
authors.forename: Elena
authors.initials: E
authors.lastname: Korchagina
authors.affiliation: Immucor, Norcross, Georgia.
authors.forename: Tom
authors.initials: T
authors.lastname: Frame
authors.affiliation: Shemyakin Institute of Bioorganic Chemistry, Moscow, Russian Federation.
authors.forename: Ivan
authors.initials: I
authors.lastname: Ryzhov
authors.affiliation: Shemyakin Institute of Bioorganic Chemistry, Moscow, Russian Federation.
authors.forename: Nicolai
authors.initials: N
authors.lastname: Bovin
authors.affiliation: Biotech Innovation Centre, Faculty of Design and Creative Technologies, AUT University, Auckland, New Zealand.
authors.forename: Stephen
authors.initials: S
authors.lastname: Henry
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry* / Q000276:immunology; D000918:Antibody Specificity; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D002460:Cell Line; D003429:Cross Reactions; D006801:Humans; D007983:Lewis Blood Group Antigens / Q000097:blood* / Q000276:immunology; D051379:Mice
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D007983:Lewis Blood Group Antigens