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Chapter 4,739

Analysis of cell surface molecules on human platelets with monoclonal antibodies identification of 4 platelet specific cell surface molecules

Hayashi, K.; Furukawa, K.

Journal of the Nagoya Medical Association 106(3): 171-179

1984


Accession: 004738397

Clones (18) of monoclonal antibodies (HPL1-18) reactive to human platelets were raised. In serological analysis using an array of human cells as targets, all the clones were proved to react specifically with cell surface antigens of human platelets. In immunoprecipitation analysis to determine the MW of recognized antigens, 4 clones (HPL1-4) precipitated glycoprotein molecules (MW 104 kdalton and 127 kdalton with 125I labeled antigens, and 92 kdalton and 122 kdalton with 3H-NaB H4 labeled antigens). They did not react with platelets obtained from Glanzmann's thrombasthenia patients. Twelve clones (HPL7-18) also precipitated glycoprotein (MW 124 kdalton with 125I labeled antigens and 132 kdalton with 3H-NaB H4 labeled antigens). These clones did not react with platelets obtained from patients with Bernard-Soulier syndrome. Evidently, HPL1-4 clones are reactive to GP IIb-IIIa and HPL7-18 clones are reactive to GPIb. HPL5 precipitated glycoprotein (MW 78 kdalton with 125I labeled antigen and 80 kdalton with 3H-NaB H4 labeled antigens) and HPL6 precipitated molecules whose MW was 151 kdalton with 125I labeled antigens. The molecules that precipitated with HPL6, however, were not labeled with 3H-NaB H4, indicating that the detected antigen molecules contained little or no sugar portion. Use of fluoroscinated monoclonal antibodies in flow cytometry was extremely useful in analyzing molecular or size abnormality of diseased platelets.

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