Utility of frozen platelets for a platelet antibody assay using flow cytometric analysis.

Diagnostic and clinical immunology Pub Date : 1988-01-01
J Lazarchick, P C Das, T J Jones, R J Russell, S A Hall
{"title":"Utility of frozen platelets for a platelet antibody assay using flow cytometric analysis.","authors":"J Lazarchick,&nbsp;P C Das,&nbsp;T J Jones,&nbsp;R J Russell,&nbsp;S A Hall","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>The immunoreactivity of a PLA1 antibody-containing serum to frozen/thawed platelets prepared by four different procedures was measured to assess the practicality of preparing a platelet reagent that could be easily stored and readily used in an immunofluorescent platelet antibody detection system. These frozen aliquots were analyzed at intervals up to 23 weeks after initial freezing and storage. For analysis, the frozen platelets were thawed, washed, resuspended to 2.5 x 10(5) platelets/mL and incubated with a dilution of fresh autologous serum that lacked platelet antibody or with a similar dilution of frozen human serum containing IgG anti-PLA1 antibody. At each interval, a freshly drawn platelet sample from the same donor was incubated in the same manner and used for comparison. All platelet mixtures were then washed and incubated with fluorescein conjugated goat F (ab')2 antihuman IgG. After repeat washing, each mixture was then analyzed with a flow cytometer for the extent of fluorescent antibody bound to each platelet mixture. Anti-PLA1 antibody reactivity with either the fresh or the frozen/thawed platelets remained stable over the period of analysis, with no significant difference in immunofluorescence with either fresh or frozen platelets as target cells. Platelet recovery following the thawing step ranged from 12% to 36% and was independent of the storage time. These studies suggest that frozen platelets can be readily used as reagents for platelet antibody assays.</p>","PeriodicalId":77705,"journal":{"name":"Diagnostic and clinical immunology","volume":"5 6","pages":"338-43"},"PeriodicalIF":0.0000,"publicationDate":"1988-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Diagnostic and clinical immunology","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

The immunoreactivity of a PLA1 antibody-containing serum to frozen/thawed platelets prepared by four different procedures was measured to assess the practicality of preparing a platelet reagent that could be easily stored and readily used in an immunofluorescent platelet antibody detection system. These frozen aliquots were analyzed at intervals up to 23 weeks after initial freezing and storage. For analysis, the frozen platelets were thawed, washed, resuspended to 2.5 x 10(5) platelets/mL and incubated with a dilution of fresh autologous serum that lacked platelet antibody or with a similar dilution of frozen human serum containing IgG anti-PLA1 antibody. At each interval, a freshly drawn platelet sample from the same donor was incubated in the same manner and used for comparison. All platelet mixtures were then washed and incubated with fluorescein conjugated goat F (ab')2 antihuman IgG. After repeat washing, each mixture was then analyzed with a flow cytometer for the extent of fluorescent antibody bound to each platelet mixture. Anti-PLA1 antibody reactivity with either the fresh or the frozen/thawed platelets remained stable over the period of analysis, with no significant difference in immunofluorescence with either fresh or frozen platelets as target cells. Platelet recovery following the thawing step ranged from 12% to 36% and was independent of the storage time. These studies suggest that frozen platelets can be readily used as reagents for platelet antibody assays.

利用流式细胞术分析,冷冻血小板用于血小板抗体测定。
测定含PLA1抗体的血清对四种不同方法制备的冷冻/解冻血小板的免疫反应性,以评估制备易于储存和易于用于免疫荧光血小板抗体检测系统的血小板试剂的实用性。在初次冷冻和储存后的23周内,对这些冷冻的等分液进行分析。将冷冻的血小板解冻、洗涤、重悬至2.5 × 10(5)个血小板/mL,并与缺乏血小板抗体的新鲜自体血清或含有IgG抗pla1抗体的类似稀释的冷冻人血清孵育。每隔一段时间,从同一供体新鲜抽取的血小板样本以同样的方式孵育并用于比较。然后清洗所有血小板混合物,并用荧光素偶联的山羊F (ab’)2抗人IgG孵育。重复洗涤后,用流式细胞仪分析每种血小板混合物的荧光抗体结合程度。在分析期间,新鲜或冷冻/解冻血小板的抗pla1抗体反应性保持稳定,新鲜或冷冻血小板作为靶细胞的免疫荧光无显著差异。解冻后的血小板回收率在12% ~ 36%之间,与保存时间无关。这些研究表明,冷冻血小板可以很容易地用作血小板抗体测定的试剂。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信