Slot blotting and flow cytometry: two efficient assays for platelet antibody screening among patients with platelet refractoriness

Vox Sanguinis ◽  
2020 ◽  
Author(s):  
Zahra Ameri ◽  
Reza Vahidi ◽  
Morteza Khaleghi ◽  
Tania Dehesh ◽  
Amir Sheikhbardsiri ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Antibody Screening ◽  
Platelet Antibody

Incidence of HPA-9b in Testing for Neonatal Alloimmune Thrombocytopenia.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3214-3214
Author(s):  
James W. Smith ◽  
Kristen Chamberlain ◽  
Diana Moffatt ◽  
Aurelio Santos ◽  
Denise Neutel ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Normal Control ◽  
Gene Frequency ◽  
Platelet Reactivity ◽  
Low Frequency ◽  
Control Population ◽  
Antigen Capture ◽  
Increased Risk ◽  
Alloimmune Thrombocytopenia ◽  
Platelet Antibody

Abstract Background: Neonatal alloimmune thrombocytopenia (NAT) is caused by maternal alloantibodies that cross the placenta and cause increased clearance of fetal platelets. The alloantibodies are directed to incompatible human platelet antigens (HPA) inherited from the father and expressed on fetal platelet glycoproteins. Most cases of NAT are caused by HPA-1a, -5a, -5b, -15a, or -15b. Some are associated with immunization to a low frequency antigen found only in the paternal lineage. We investigated the incidence of the low frequency antigen HPA-9b in a population with suspected NAT, and compared the frequency to that found in a normal control population. HPA-9b is caused by a Val837Met polymorphism on alpha-IIb and is expressed only in a rare subset of individuals with a corresponding serine at position 843 that defines the HPA-3b antigen. Methods: We used PCR-SSP for HPA-9b genotyping. Antibody studies were performed by incubating maternal serum with intact platelets using flow cytometry, monoclonal antigen capture enzyme immunoassay, and radioimmunoprecipitation. Results: Of 294 normal control blood donors, 4 typed positive for HPA-9b, yielding a gene frequency of 0.0068. However, if only HPA-3b positive donor controls are considered (n=177) the gene frequency for HPA-9b is 0.011. Archived DNA from HPA-3b positive (HPA-3a3b or -3b3b) fathers of children with NAT were tested for HPA-9b. Of 94 fathers, 3 (3.2%) typed HPA-9a9b, yielding a gene frequency for HPA-9b of 0.016, not significantly different from the frequency for HPA-9b found in the HPA-3b-positive healthy controls (p>0.1). Similarily, when DNA from HPA-3b positive mothers was tested in tandem, 3 of 60 were positive for HPA-9b, yielding a gene frequency of 0.026. Of the 3 cases of suspected NAT (platelet count range 15,000 to 63,000) in which the mother and father were incompatible for HPA-9b, 2 had no other platelet antigen incompatibility, and one was incompatible for HPA-15b. None of the 3 maternal sera was positive for anti-platelet reactivity using radioimmunoprecipitation and paternal platelets as a target, and no anti-platelet antibody was detected either by flow cytometry or by GPIIb/IIIa antigen capture immunoassay with platelets from a HPA-9a9b donor. Interpretation: These results support the findings that HPA-9b is distributed in the general population at a frequency higher than initially suspected. However, we also found that the frequency of HPA-9b in fathers of children with suspected NAT was similar to that in the normal control population when only HPA-3b-positive (HPA-3a3b or -3b3b) targets were analyzed. Therefore, in the absence of a positive anti-platelet antibody, the significance of a maternal/paternal incompatibility for HPA-9b alone is not predictive of an increased risk of NAT.

scholarly journals POSSIBLE METHOD FOR IRREGULAR ANTIBODY SCREENING BY FLOW CYTOMETRY

2019 ◽  
Vol 65 (5) ◽  
pp. 782-792
Author(s):  
Takayuki Maruhashi ◽  
Yousuke Kitazume ◽  
Kana Tashiro ◽  
Izuru Nagaoka ◽  
Sayaka Takeuchi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Antibody Screening

Relationship between CDC cross-match in liver recipients and antibody screening by flow cytometry

2003 ◽  
Vol 35 (5) ◽  
pp. 1894-1895 ◽  
Author(s):  
M Muro ◽  
F Sánchez-Bueno ◽  
L Marín ◽  
A Torío ◽  
M.R Moya-Quiles ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Antibody Screening ◽  
Cross Match

scholarly journals Application of High-Throughput Flow Cytometry in Early Drug Discovery: An AstraZeneca Perspective

2018 ◽  
Vol 23 (7) ◽  
pp. 719-731
Author(s):  
Mei Ding ◽  
Roger Clark ◽  
Catherine Bardelle ◽  
Anna Backmark ◽  
Tyrrell Norris ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Single Cells ◽  
Phenotypic Screening ◽  
Antibody Screening ◽  
Structure Activity ◽  
High Level ◽  
Early Drug

Flow cytometry is a powerful tool providing multiparametric analysis of single cells or particles. The introduction of faster plate-based sampling technologies on flow cytometers has transformed the technology into one that has become attractive for higher throughput drug discovery screening. This article describes AstraZeneca’s perspectives on the deployment and application of high-throughput flow cytometry (HTFC) platforms for small-molecule high-throughput screening (HTS), structure–activity relationship (SAR) and phenotypic screening, and antibody screening. We describe the overarching HTFC workflow, including the associated automation and data analysis, along with a high-level overview of our HTFC assay portfolio. We go on to discuss the practical challenges encountered and solutions adopted in the course of our deployment of HTFC, as well as future enhancements and expansion of the technology to new areas of drug discovery.

Comparison of Antibody Screening Methods Using Flow Cytometry With Cells or Immobilized Purified HLA Antigens

1998 ◽  
Vol 65 (Supplement) ◽  
pp. 145
Author(s):  
Rhonda Senkbeil ◽  
James Shoaf ◽  
Charyl Rowe ◽  
Stephanie Tyler ◽  
Mark Deierhoi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Hla Antigens ◽  
Screening Methods ◽  
Antibody Screening

scholarly journals Platelet antibody detection by flow cytometry: an effective method to evaluate and give transfusional support in platelet refractoriness

2013 ◽  
Vol 35 (4) ◽  
Author(s):  
Carolina Bonet Bub ◽  
Beatriz Moraes Martinelli ◽  
Thayná Mendonça Avelino ◽  
Ana Cláudia Gonçalez ◽  
Maria de Lourdes Barjas-Castro ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Antibody Detection ◽  
Platelet Antibody

Antenatal platelet antibody testing by flow cytometry-results of a pilot study

Transfusion ◽  
1994 ◽  
Vol 34 (11) ◽  
pp. 961-965 ◽  
Author(s):  
LR Marshall ◽  
FE Brogden ◽  
TS Roper ◽  
AL Barr
Keyword(s):  
Flow Cytometry ◽  
Pilot Study ◽  
Antibody Testing ◽  
Platelet Antibody
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