PMID- 24785939
OWN - NLM
STAT- MEDLINE
DCOM- 20150330
LR  - 20191210
IS  - 1399-0039 (Electronic)
IS  - 0001-2815 (Linking)
VI  - 84
IP  - 2
DP  - 2014 Aug
TI  - Evaluation of a competitive enzyme-linked immunosorbent assay for measurements of 
      soluble HLA-G protein.
PG  - 206-15
LID - 10.1111/tan.12357 [doi]
AB  - The human leukocyte antigen (HLA) class Ib molecule, HLA-G, has gained increased 
      attention because of its assumed important role in immune regulation. The HLA-G 
      protein exists in several soluble isoforms. Most important are the actively 
      secreted HLA-G5 full-length isoform generated by alternative splicing retaining 
      intron 4 with a premature stop codon, and the cleavage of full-length 
      membrane-bound HLA-G1 from the cell surface, so-called soluble HLA-G1 (sHLA-G1). 
      A specific and sensitive immunoassay for measurements of soluble HLA-G is 
      mandatory for conceivable routine testing and research projects. We report a 
      novel method, a competitive immunoassay, for measuring HLA-G5/sHLA-G1 in 
      biological fluids. The sHLA-G immunoassay is based upon a competitive 
      enzyme-linked immunosorbent assay (ELISA) principle. It includes a recombinant 
      sHLA-G1 protein in complex with beta2-microglobulin and a peptide as a standard, 
      biotinylated recombinant sHLA-G1 as an indicator, and the MEM-G/9 anti-HLA-G 
      monoclonal antibody (mAb) as the capture antibody. The specificity and 
      sensitivity of the assay were evaluated. Testing with different recombinant HLA 
      class I proteins and different anti-HLA class I mAbs showed that the sHLA-G 
      immunoassay was highly specific. Optimal combinations of competitor sHLA-G1 and 
      capture mAb concentrations were determined. Two versions of the assay were 
      tested. One with a relatively wide dynamic range from 3.1 to 100.0 ng/ml, and 
      another more sensitive version ranging from 1.6 to 12.5 ng/ml. An intra-assay 
      coefficient of variation (CV) of 15.5% at 88 ng/ml and an inter-assay CV of 23.1% 
      at 39 ng/ml were determined. An assay based on the competitive sHLA-G ELISA may 
      be important for measurements of sHLA-G proteins in several conditions: assisted 
      reproduction, organ transplantation, cancer, and certain pregnancy complications, 
      both in research studies and possibly in the future also for clinical routine 
      use.
CI  - (c) 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
FAU - Rasmussen, M
AU  - Rasmussen M
AD  - Department of Clinical Biochemistry, Centre for Immune Regulation and 
      Reproductive Immunology (CIRRI), Copenhagen University Hospital (Roskilde) and 
      Roskilde Hospital, Roskilde, Denmark.
FAU - Dahl, M
AU  - Dahl M
FAU - Buus, S
AU  - Buus S
FAU - Djurisic, S
AU  - Djurisic S
FAU - Ohlsson, J
AU  - Ohlsson J
FAU - Hviid, T V F
AU  - Hviid TV
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140502
PL  - England
TA  - Tissue Antigens
JT  - Tissue antigens
JID - 0331072
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Culture Media, Conditioned)
RN  - 0 (HLA-G Antigens)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Antibodies, Monoclonal/immunology
MH  - Antibody Specificity/immunology
MH  - Biotinylation
MH  - Cell Line, Tumor
MH  - Culture Media, Conditioned/chemistry
MH  - Enzyme-Linked Immunosorbent Assay/*methods
MH  - HLA-G Antigens/*blood/immunology
MH  - Humans
MH  - Recombinant Proteins/immunology
MH  - Reference Standards
MH  - Sensitivity and Specificity
MH  - Solubility
OTO - NOTNLM
OT  - competitive enzyme-linked immunosorbent assay
OT  - human leukocyte antigen
OT  - human leukocyte antigen class Ib
OT  - soluble human leukocyte antigen-G
EDAT- 2014/05/03 06:00
MHDA- 2015/03/31 06:00
CRDT- 2014/05/03 06:00
PHST- 2013/10/28 00:00 [received]
PHST- 2014/03/24 00:00 [revised]
PHST- 2014/03/25 00:00 [accepted]
PHST- 2014/05/03 06:00 [entrez]
PHST- 2014/05/03 06:00 [pubmed]
PHST- 2015/03/31 06:00 [medline]
AID - 10.1111/tan.12357 [doi]
PST - ppublish
SO  - Tissue Antigens. 2014 Aug;84(2):206-15. doi: 10.1111/tan.12357. Epub 2014 May 2.