title: Antibodies to biotinylated red blood cells in adults and infants: improved detection, partial characterization, and dependence on red blood cell-biotin dose.
abstract: Biotin-labeled red blood cells (BioRBCs) are used for in vivo kinetic studies. Because BioRBC dosing occasionally induces antibodies, a sensitive and specific anti-BioRBC detection assay is needed.
  
  
  
  Aims were to 1) develop a gel card assay to evaluate existing, naturally occurring and BioRBC-induced plasma antibodies, 2) compare gel card and tube agglutination detection results, and 3) test for a relationship of antibody induction and BioRBC dose. Reagent BioRBCs were prepared using sulfo-NHS biotin ranging from densities 18 (BioRBC-18) to 1458 (BioRBC-1458) µg/mL RBCs.
  
  
  
  Among BioRBC-exposed subjects, gel card and tube agglutination results were concordant in 21 of 22 adults and all 19 infant plasma samples. Gel card antibody detection sensitivity was more than 10-fold greater than tube agglutination. Twelve to 16 weeks after BioRBC exposure, induced anti-antibodies were detected by gel card in three of 26 adults (12%) at reagent densities BioRBC-256 or less, but in none of 41 infants. Importantly, induced anti-BioRBC antibodies were associated with higher BioRBC dose (p = 0.008); no antibodies were detected in 18 subjects who received BioRBC doses less than or equal to BioRBC-18. For noninduced BioRBC antibodies, six of 1125 naïve adults (0.3%) and none of 46 naïve infants demonstrated existing anti-BioRBC antibodies using reagent BioRBC-140 or -162. Existing anti-BioRBCs were all neutralized by biotin compounds, while induced antibodies were not.
  
  
  
  The gel card assay is more sensitive than the tube agglutination assay. We recommend reagent BioRBC-256 for identifying anti-BioRBCs. Use of a low total RBC biotin label dose (≤ BioRBC-18) may minimize antibody induction.
authors.affiliation: Department of Pediatrics, Carver College of Medicine, Iowa City, Iowa.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Schmidt
authors.affiliation: Department of Biochemistry & Molecular Biology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, Arkansas.|Department of Pediatrics, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, Arkansas.
authors.forename: Donald M
authors.initials: DM
authors.lastname: Mock
authors.affiliation: Department of Internal Medicine, Division of Hematology/Oncology, University of Cincinnati College of Medicine.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Franco
authors.affiliation: Department of Internal Medicine, Division of Endocrinology, Diabetes & Metabolism, University of Cincinnati College of Medicine and Medical Service, Cincinnati VA Medical Center, Cincinnati, Ohio.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Cohen
authors.affiliation: Cerus Corporation, Concord, California.
authors.forename: Anne K
authors.initials: AK
authors.lastname: North
authors.affiliation: Hoxworth Blood Center and the Department of Pediatrics, The University of Cincinnati, Cincinnati, Ohio.
authors.forename: José A
authors.initials: JA
authors.lastname: Cancelas
authors.affiliation: Institut für Transfusionsmedizin und Immunhämatologie, Klinikum der Goethe Universität, Frankfurt am Main, DRK-Blutspendedienst, Baden-Württemberg-Hessen, Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: Geisen
authors.affiliation: Department of Pediatrics, Carver College of Medicine, Iowa City, Iowa.|Department of Pathology, Carver College of Medicine, Iowa City, Iowa.
authors.forename: Ronald G
authors.initials: RG
authors.lastname: Strauss
authors.affiliation: Department of Intensive Care and Laboratory of Experimental Intensive Care and Anesthesia, Academic Medical Center, Amsterdam, the Netherlands.
authors.forename: Alexander P
authors.initials: AP
authors.lastname: Vlaar
authors.affiliation: Department of Pediatrics, Carver College of Medicine, Iowa City, Iowa.
authors.forename: Demet
authors.initials: D
authors.lastname: Nalbant
authors.affiliation: Department of Pediatrics, Carver College of Medicine, Iowa City, Iowa.
authors.forename: John A
authors.identifier: 0000-0001-6543-5927
authors.initials: JA
authors.lastname: Widness
mesh_terms: D000328:Adult; D000372:Agglutination Tests; D000906:Antibodies / Q000276:immunology*; D001681:Biological Assay / Q000379:methods; D001710:Biotin / Q000737:chemistry*; D019921:Biotinylation; D004912:Erythrocytes / Q000276:immunology*; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D013388:Succinimides / Q000737:chemistry
chemical_list: D000906:Antibodies; C465543:N-hydroxysulfosuccimide; D013388:Succinimides; D001710:Biotin