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PMID: 2547875 Published · ppublish English Journal Article

Purification of human blood eosinophils by negative selection using immunomagnetic beads.

Journal of immunological methods ·Vol. 122 ·No. 1 ·1989-08-15 ·Pages 97-103

Hansel TT, Pound JD, Pilling D, Kitas GD, Salmon M, Gentle TA, Lee SS, Thompson RA

Abstract

A simple method for isolating highly purified eosinophils from human blood is described. Buffy coats from normal individuals (eosinophil counts less than 0.4 x 10(9)/litre) were centrifuged through a two layer Percoll density gradient, to produce a granulocyte fraction containing neutrophils and eosinophils. Neutrophils were extracted from this fraction using a monoclonal antibody (CLB FcR gran 1) against CD 16 (Fc gamma R III) in a direct or indirect selection procedure using immunomagnetic beads (Dynabeads). This negative immunoselection produced eosinophils of greater purity and with a superior capacity to mount a respiratory burst than eosinophils isolated by a method employing metrizamide.

MeSH Terms
Antigens, Differentiation/immunology Cell Separation/methods Centrifugation, Density Gradient Eosinophils/immunology Humans Immunologic Techniques Luminescent Measurements Magnetics Metrizamide Neutrophils/immunology Povidone Receptors, Fc/immunology Receptors, IgG Silicon Dioxide
Chemicals
Antigens, Differentiation Receptors, Fc Receptors, IgG Percoll Silicon Dioxide Povidone Metrizamide
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Hansel T T
Regional Department of Immunology, East Birmingham Hospital, U.K.
Pound J D
Pilling D
Kitas G D
Salmon M
Gentle T A
Lee S S
Thompson R A
Article Info
Journal
Journal of immunological methods
Abbr.
J Immunol Methods
ISSN
0022-1759
Published
1989-08-15
Pages
97-103
Language
English
Region
Netherlands
NLM ID
1305440
Subset
IM
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