Selection and validation of antibody clones against IgG and IgA subclasses in switched memory B-cells and plasma cells
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00064203%3A_____%2F19%3A10402066" target="_blank" >RIV/00064203:_____/19:10402066 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11130/19:10402066
Result on the web
<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=lJRyy-ACRc" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=lJRyy-ACRc</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.jim.2017.09.008" target="_blank" >10.1016/j.jim.2017.09.008</a>
Alternative languages
Result language
angličtina
Original language name
Selection and validation of antibody clones against IgG and IgA subclasses in switched memory B-cells and plasma cells
Original language description
The clinical value of assessing immunoglobulin (Ig)G and IgA subclasses in addition to the isotypes of soluble Igs in serum has been well established. > 20 years ago, the International Union of Immunological Societies and the World Health Organization performed collaborative studies in order to validate antibody (Ab) clones for the detection of IgG and IgA subclasses for a broad range of laboratory assays, except for flow cytometry. Here we analyzed the performance of commercially available Ab clones to detect IgG and IgA subclasses in memory B-cells and plasma cells (PCs) by flow cytometry. In a first step, 28 Ab clones were evaluated in peripheral blood from healthy donors. Only 17/28 clones showed reactivity against IgG and IgA subclasses expressed on the B-cell and PC surface membrane, including Ab clones for IgG(1) (SAG1, HP6188, HP6001 and HP6186), IgG(2) (SAG2, HP6014 and HP6002), IgG(3) (SAG3, HP6095 and HP6050), IgG(4) (SAG4), IgA(1) (SAA1, H69-11.4 and B3506B4) and IgA(2) (SAA2, 2E2, and A9604D2). In a second step, for each Ig subclass a single clone was selected according to its specificity and fluorescence intensity (resolution power), for further more detailed validation (SAG1, SAG2, SAG3, SAG4, SAA1 and SAA2). This validation process was carried out in 4 different laboratories by testing the selected Ab clones in human peripheral blood, bone marrow and tonsil samples, using different staining protocols (e.g. surface membrane and/or cytoplasmic staining). All selected Ab clones displayed strong positivity, high specificity and optimal resolution between negative and positive cells. Alternative Ab clones were also validated. Thus, our results show the feasibility of using the validated Ig subclass Ab clones in combination with other B cell-associated markers for detailed dissection of the memory B-cell and PC compartments that express distinct Ig subclasses in different human tissues.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
30204 - Oncology
Result continuities
Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2019
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Immunological Methods
ISSN
0022-1759
e-ISSN
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Volume of the periodical
475
Issue of the periodical within the volume
SI
Country of publishing house
NL - THE KINGDOM OF THE NETHERLANDS
Number of pages
12
Pages from-to
112372
UT code for WoS article
000502791200013
EID of the result in the Scopus database
2-s2.0-85032632148