Antibodies to biotinylated red blood cells in adults and infants: improved detection, partial characterization, and dependence on red blood cell-biotin dose

Abstract

Background: 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.

Study design and methods: 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.

Results: 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.

Conclusions: 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.

Conflict of interest statement

Conflict of Interest Statements:

Robert L. Schmidt: None.

Donald M Mock:

1. Consultant Medday Pharmaceuticals

Robert S. Franco: None.

Robert M. Cohen: None.

Anne K. North:

1. Employee Cerus Corporation, 2550 Stanwell Drive, Concord CA 94520, USA

Jose A. Cancelas:

1. None for companies. Research support from NIH, US Dept. of Defense, Leukemia & Lymphoma Society of North America, William & Blanche Lawrence Hughes Foundation, Cerus Co., Terumo BCT, Zimmet Biomed., Cellphyre, Inc., and New Health Sciences, Inc.

2. Unpaid advisory committee New Health Sciences, Inc.

Christof Geisen: None.

Ronald G. Strauss: None.

Alexander P. Vlaar: None.

Demet Nalbant: None.

John A. Widness:

1. Consultant HemoGenix (http://hemogenix.com)

2. Loan agreement Sysmex America, Inc., Hematology Analyzer

© 2017 AABB.

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