Production of human monoclonal anti‐Jk3, recognising an epitope including the Jk<sup>a</sup>/Jk<sup>b</sup> polymorphic site of the Kidd glycoprotein

Toyoda, Chizu; Suzuki, Yumi; Tsuneyama, Hatsue; Onodera, T.; Masuno, Atsuko; Yabe, Ryuichi; Ogasawara, Kenichi; Okuda, Makoto; Nakajima, Kazunori; Uchikawa, Makoto

doi:10.1111/tme.12146

Cited by 7 publications

(7 citation statements)

References 23 publications

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“…Genomic DNA was extracted from whole blood using the QIAamp DNA Blood Mini Kit (Qiagen, Chuo‐ku, Tokyo, Japan) in accordance with the manufacturer's instructions. Reticulocyte RNA was extracted and cDNA was synthesized in accordance with the methods described previously .…”

Section: Methodsmentioning

confidence: 99%

Weak D alleles in Japanese: a c.960G>A silent mutation in exon 7 of the RHD gene that affects D expression

et al. 2015

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Section: Methodsmentioning

confidence: 99%

Weak D alleles in Japanese: a c.960G>A silent mutation in exon 7 of the RHD gene that affects D expression

et al. 2015

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“…Hybridomas were selected by an ouabain-containing HAT medium. Single clones were isolated by limiting dilution [ 30 ]. Antibodies secreted by hybridomas were detected by a hemagglutination assay with known GP.Mur RBCs.…”

Section: Methodsmentioning

confidence: 99%

Universal Detection of Mia Antigen and Frequencies of Glycophorin Hybrids among Blood Donors in Taiwan by Human Monoclonal Antibodies against Mia (MNS7), Mur (MNS10), and MUT (MNS35) Antigens

et al. 2021

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Glycophorin hybrids such as GP.Mur are common in Southeast Asians. In Taiwan, clinically significant alloantibodies to the GP.Mur phenotype are the most important issue in blood banks. A large-scale screening of glycophorin hybrids in the Taiwanese population is urgently needed to ensure transfusion safety. Four clones of human hybridomas that secrete anti-Mia, anti-MUT, and anti-Mur were established by fusing human B-lymphocytes and myeloma cells (JMS-3). The specificity of each monoclonal antibody (MoAb) was characterized. Three MoAbs were applied on an Automated Pretransfusion Blood Testing Analyzer (PK7300/PK7400) for donor screening. Genotyping was performed to determine the detailed subgrouping of glycophorin hybrids. Four MoAbs are IgM antibodies. Anti-Mia (377T) binds to 46DXHKRDTYA54, 48HKRDTYAAHT57 peptides, and anti-Mia (367T) binds to 43QTNDXHKRD51 peptides (X indicates T, M, or K). Anti-Mur is reactive with 49KRDTYPAHTA58 peptides. Anti-MUT is reactive with 47KHKRDTYA54. A total of 78,327 donors were screened using three MoAbs, and 3690 (4.71%) were GP.Mur, 20 (0.025%) were GP.Hut, and 18 (0.022%) were GP.Vw. When the Mia antigen was introduced as routine screening, the frequency of Mi(a+) among blood donors in Taiwan was 4.66% (67,348/1,444,541). Mia antigen was implemented as a routine blood testing, and the results were labeled on all red blood cell (RBC) units.

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“…Flow cytometric analysis of S and s antigens on RBCs was performed according to the method described previously [7] using anti‐S (Bioclone, Ortho Clinical Diagnostics) and anti‐s (Bioclone, Ortho Clinical Diagnostics) as the primary antibodies, and R‐phycoerythrin‐conjugated anti‐human IgG goat antibody (PROTOS Immunoresearch, Burlingame, CA, USA) as the second antibody. Common RBCs were used as controls, and the relative mean fluorescence intensities (MFIs) were compared.…”

Section: Methodsmentioning

confidence: 99%

An unusual variant glycophorin expressing protease‐resistant M antigen encoded by the GYPB‐E(2‐4)‐B hybrid gene

et al. 2020

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Background and objectives MNS is a highly polymorphic blood group comprising 49 antigens recognized by International Society of Blood Transfusion, some of which may have been generated by genomic recombination among the closely linked genes GYPA, GYPB and GYPE. The GYPE gene has an almost identical sequence to GYPA*01 allele in exon 2 (99% homology), which accounts for M antigen. We investigated an unusual glycophorin molecule with protease-resistant M antigen. Methods Blood samples were screened by an automated blood typing system (PK7300) using bromelain-treated red blood cells (RBCs) and murine monoclonal anti-M. The M-positive RBC samples were analysed by immunoblotting using anti-M as the primary antibody. GYPA, GYPB and GYPE genes were analysed by polymerase chain reaction (PCR), cloning and sequencing using reticulocyte mRNA and genomic DNA. Results Serological tests and immunoblotting revealed that 103 of the 193 009 individuals (0Á0534%) expressed protease-resistant M-active glycophorin having a molecular weight of 20 kDa. All the 103 individuals were S+ s-or S-s+. When reticulocyte mRNA from the individuals with M-active glycophorin (20 kDa) was examined by PCR and cloning followed by sequencing, a novel GYPE-B hybrid transcript was identified. Long-range PCR and sequencing using genomic DNA revealed that the individuals had a GYPB-E(2-4)-B hybrid gene. This hybrid gene was predicted to encode a 59-amino-acid mature glycoprotein that expresses no S or s antigens Conclusions The prevalence of the M-active glycophorin (20 kDa) in the Japanese population is 0Á0534%. This glycophorin is predicted to be a 59 amino acids polypeptide encoded by the novel GYPB-E(2-4)-B hybrid gene.

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Production of human monoclonal anti‐Jk3, recognising an epitope including the Jk^a/Jk^b polymorphic site of the Kidd glycoprotein

Abstract: We established a human hybridoma cell line secreting monoclonal anti-Jk3 (HIRO-294). This antibody had unique specificity, recognising the Kidd glycoprotein including the Jk(a) /Jk(b) polymorphic site.

Cited by 7 publications

References 23 publications

Weak D alleles in Japanese: a c.960G>A silent mutation in exon 7 of the RHD gene that affects D expression

Weak D alleles in Japanese: a c.960G>A silent mutation in exon 7 of the RHD gene that affects D expression

Universal Detection of Mia Antigen and Frequencies of Glycophorin Hybrids among Blood Donors in Taiwan by Human Monoclonal Antibodies against Mia (MNS7), Mur (MNS10), and MUT (MNS35) Antigens

An unusual variant glycophorin expressing protease‐resistant M antigen encoded by the GYPB‐E(2‐4)‐B hybrid gene

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Production of human monoclonal anti‐Jk3, recognising an epitope including the Jka/Jkb polymorphic site of the Kidd glycoprotein

Abstract: We established a human hybridoma cell line secreting monoclonal anti-Jk3 (HIRO-294). This antibody had unique specificity, recognising the Kidd glycoprotein including the Jk(a) /Jk(b) polymorphic site.

Cited by 7 publications

References 23 publications

Weak D alleles in Japanese: a c.960G>A silent mutation in exon 7 of the RHD gene that affects D expression

Weak D alleles in Japanese: a c.960G>A silent mutation in exon 7 of the RHD gene that affects D expression

Universal Detection of Mia Antigen and Frequencies of Glycophorin Hybrids among Blood Donors in Taiwan by Human Monoclonal Antibodies against Mia (MNS7), Mur (MNS10), and MUT (MNS35) Antigens

An unusual variant glycophorin expressing protease‐resistant M antigen encoded by the GYPB‐E(2‐4)‐B hybrid gene

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Production of human monoclonal anti‐Jk3, recognising an epitope including the Jk^a/Jk^b polymorphic site of the Kidd glycoprotein