A new method for histamine release from purified peripheral blood basophils using monoclonal antibody-coated magnetic beads.
A new method for evaluating histamine release from purified basophils was developed. Basophil-containing leukocytes were directly purified from a small amount of peripheral blood using monoclonal antibody BA312-coated magnetic beads. The purified basophils still rosetted to magnetic beads maintained a normal response to anti-IgE and to dust mite allergen in comparison with the conventional method using washed leukocytes. This methodology facilitates the purification of basophils, anti-IgE- and allergen-induced histamine release, and subsequent histamine determination within only 3 h. The released histamine was analyzed by an enzyme-linked immunosorbent assay (ELISA) with a characteristic detection profile. Since all steps were performed in 96-well microplates, many clinical samples could be analyzed at the same time, permitting easy applications in routine laboratories.
Nishi
H
H
Diagnostic Science Division, Shionogi and Co., Ltd., 2-5-1 Mishima, Settsu-shi, Osaka, Japan. hiroshi.nishi@shionogi.co.jp
Nishimura
S
S
Higashiura
M
M
Ikeya
N
N
Ohta
H
H
Tsuji
T
T
Nishimura
M
M
Ohnishi
S
S
Higashi
H
H
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001491:Basophils / Q000166:cytology / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006632:Histamine / Q000032:analysis; D006636:Histamine Release / Q000276:immunology*; D006801:Humans; D006967:Hypersensitivity / Q000175:diagnosis*; D018189:Immunomagnetic Separation / Q000379:methods*
D000911:Antibodies, Monoclonal; D006632:Histamine