A novel series of anti-human glycophorin A (CD235a) antibodies defining five extra- and intracellular epitopes

Karsten, Uwe; Butschak, G; Stahn, Renate; Goletz, Steffen

doi:10.1016/j.intimp.2010.08.001

Cited by 12 publications

(6 citation statements)

References 38 publications

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“…These affinities rival those of most naturally occurring lectins, as well as conventional and recombinant antibodies. For comparison, a multimeric anti-TFα scFv had dissociation constants for aGPA of 88 and 220 nM, for the tetrameric and trimeric forms of the antibody, respectively; the antiviral lectin griffithsin binds carbohydrates of gp120 with a dissociation constant of 8 nM; and a pentameric IgM antibody was isolated with an exceptionally high affinity constant of 200 pM for a glycopeptide epitope in aGPA . For lambody clones that are isolated with only moderate affinity for glycans, it should be possible to improve the binding by in vitro affinity maturation of the VLRs, as previously reported for VLR clones reactive to the protein lysozyme, with 100-fold improvement following a single cycle of random mutagenesis and 1,300-fold improvement following targeted mutagenesis …”

Section: Resultsmentioning

confidence: 99%

Sugar-Binding Proteins from Fish: Selection of High Affinity “Lambodies” That Recognize Biomedically Relevant Glycans

et al. 2012

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Glycan-binding proteins are important for a wide variety of basic research and clinical applications, but proteins with high affnity and selectivity for carbohydrates are diffcult to obtain. Here we describe a facile and cost-effective strategy to generate monoclonal lamprey antibodies, called lambodies, that target glycan determinants. We screened a library of yeast surface-displayed (YSD) lamprey variable lymphocyte receptors (VLR) for clones that can selectively bind various biomedically important glycotopes. These glycoconjugates included tumor-associated carbohydrate antigens (Tn and TFα), Lewis antigens (LeA and LeX), N-glycolylneuraminic acid, targets of broadly neutralizing HIV antibodies (poly-Man9 and the HIV gp120), and the glycoproteins asialo-ovine submaxillary mucin (aOSM) and asialo-human glycophorin A (aGPA). We isolated clones that bind each of these targets in a glycan-dependent manner and with very strong binding constants, for example, 6.2 nM for Man9 and 44.7 nM for gp120, determined by surface plasmon resonance (SPR). One particular lambody, VLRB.aGPA.23, was shown by glycan array analysis to be selective for the blood group H type 3 trisaccharide (BG-H3, Fucα1-2Galβ1-3GalNAcα), aGPA, and TFα (Galβ1-3GalNAcα), with affnity constants of 0.2, 1, and 8 nM, respectively. In human tissue microarrays this lambody selectively detected cancer-associated carbohydrate antigens in 14 different types of cancers. It stained 27% of non-small cell lung cancer (NSCLC) samples in a pattern that correlated with poor patient survival. Lambodies with exquisite affnity and selectivity for glycans may find myriad uses in glycobiology and biomedical research.

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Section: Resultsmentioning

confidence: 99%

Sugar-Binding Proteins from Fish: Selection of High Affinity “Lambodies” That Recognize Biomedically Relevant Glycans

et al. 2012

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“…We evaluated expression of several markers at different time points to determine the differentiation progress of each cell line. We measured the expression of CD235 (Glycophorin A), a hallmark of erythroid-specific lineage 42 . We detected the expression of CD235 by flow cytometry in all samples at the end of the differentiation regime (day 7).…”

Section: Resultsmentioning

confidence: 99%

The miR-144/Hmgn2 regulatory axis orchestrates chromatin organization during erythropoiesis

Kretov,

Folkes,

Mora-Martin

et al. 2024

Nat Commun

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Differentiation of stem and progenitor cells is a highly regulated process that involves the coordinated action of multiple layers of regulation. Here we show how the post-transcriptional regulatory layer instructs the level of chromatin regulation via miR-144 and its targets to orchestrate chromatin condensation during erythropoiesis. The loss of miR-144 leads to impaired chromatin condensation during erythrocyte maturation. Among the several targets of miR-144 that influence chromatin organization, the miR-144-dependent regulation of Hmgn2 is conserved from fish to humans. Our genetic probing of the miR-144/Hmgn2 regulatory axis establish that intact miR-144 target sites in the Hmgn2 3’UTR are necessary for the proper maturation of erythrocytes in both zebrafish and human iPSC-derived erythroid cells while loss of Hmgn2 rescues in part the miR-144 null phenotype. Altogether, our results uncover miR-144 and its target Hmgn2 as the backbone of the genetic regulatory circuit that controls the terminal differentiation of erythrocytes in vertebrates.

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“…CD235a marks the transmembrane glycoprotein glycophorin A (GPA), its presence is restricted to erythrocytes and their precursors. It is responsible for the shape of red blood cells [56].…”

Section: Discussionmentioning

confidence: 99%

An Alternative Application of Magnetic-Activated Cell Sorting: CD45 and CD235a Based Purification of Semen and Testicular Tissue Samples

Czétány,

Balló,

Márk

et al. 2024

IJMS

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Magnetic activated cell sorting (MACS) is a well-known sperm selection technique, which is able to remove apoptotic spermatozoa from semen samples using the classic annexinV based method. Leukocytes and erythrocytes in semen samples or in testicular tissue processed for in vitro fertilization (IVF) could exert detrimental effects on sperm. In the current study, we rethought the aforementioned technique and used magnetic microbeads conjugated with anti-CD45/CD235a antibodies to eliminate contaminating leukocytes and erythrocytes from leukocytospermic semen samples and testicular tissue samples gained via testicular sperm extraction (TESE). With this technique, a 15.7- and a 30.8-fold reduction could be achieved in the ratio of leukocytes in semen and in the number of erythrocytes in TESE samples, respectively. Our results show that MACS is a method worth to reconsider, with more potential alternative applications. Investigations to find molecules labeling high-quality sperm population and the development of positive selection procedures based on these might be a direction of future research.

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A novel series of anti-human glycophorin A (CD235a) antibodies defining five extra- and intracellular epitopes

Cited by 12 publications

References 38 publications

Sugar-Binding Proteins from Fish: Selection of High Affinity “Lambodies” That Recognize Biomedically Relevant Glycans

Sugar-Binding Proteins from Fish: Selection of High Affinity “Lambodies” That Recognize Biomedically Relevant Glycans

The miR-144/Hmgn2 regulatory axis orchestrates chromatin organization during erythropoiesis

An Alternative Application of Magnetic-Activated Cell Sorting: CD45 and CD235a Based Purification of Semen and Testicular Tissue Samples

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