Anti-Human Embryonic Stem Cell Monoclonal Antibody Hesca-2 Binds to a Glycan Epitope Commonly Found on Carcinomas

Shoreibah, Mohamed; Jackson, Crystal Lyles; Price, Paul W.; Meagher, Richard B.; Godwin, Andrew K.; Cai, Qi; Gildersleeve, Jeffrey C.

doi:10.1089/scd.2010.0172

Cited by 8 publications

(7 citation statements)

References 53 publications

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“…Of particular relevance to this study, many of the array components contain glycan sequences that are closely related to GD2 such as GD1a, GD1b, GD3, GM1a, GM1b, GM2, GM3, GT1a, GT2, GT3, GQ2, and GA2 (Figure 1). We (Gildersleeve et al, 2008; Gildersleeve and Wright, 2016; Oyelaran et al, 2009; Shoreibah et al, 2011; Zhang et al, 2010) and others (Chang et al, 2010; Dupin et al, 2015; Godula and Bertozzi, 2012; Goudot et al, 2013; Hung et al, 2013; Jaipuri et al, 2008; Karamanska et al, 2008; Ligeour et al, 2015; Park and Shin, 2007; Shivatare et al, 2013; Song et al, 2008; Wang et al, 2008) have used glycan microarrays to evaluate apparent K D values for carbohydrate-protein interactions and have found good agreement with previously published values measured by surface plasmon resonance.…”

Section: Resultssupporting

confidence: 81%

Therapeutic Antibodies to Ganglioside GD2 Evolved from Highly Selective Germline Antibodies

et al. 2017

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Summary Antibodies play a crucial role in host defense and are indispensable research tools, diagnostics, and therapeutics. Antibody generation involves binding of genomically encoded germline antibodies followed by somatic hypermutation and in vivo selection to obtain antibodies with high affinity and selectivity. Understanding this process is critical for developing monoclonal antibodies, designing effective vaccines, and understanding autoantibody formation. Prior studies have found that antibodies to haptens, peptides, and proteins evolve from polyspecific germline antibodies. The immunological evolution of antibodies to mammalian glycans has not been studied. Using glycan microarrays, protein microarrays, cell binding studies, and molecular modeling, we demonstrate that therapeutic antibodies to the tumor-associated ganglioside GD2 evolved from highly specific germline precursors. The results have important implications for developing vaccines and monoclonal antibodies that target carbohydrate antigens. In addition, they demonstrate an alternative pathway for antibody evolution within the immune system that is distinct from the polyspecific germline pathway.

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Section: Resultssupporting

confidence: 81%

Therapeutic Antibodies to Ganglioside GD2 Evolved from Highly Selective Germline Antibodies

et al. 2017

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“…Alternatively, IgM antibodies may have higher overall binding avidity toward glycan antigens than IgG and IgA. In other studies we have found that monoclonal IgM antibodies can bind very tightly to glycans on our array surface, with apparent Kd values in the low nanomolar to subnanomolar range [ 74 , 75 ]. Additionally, IgM may bind faster than IgG and the experimental conditions may not allow sufficient time for full equilibration.…”

Section: Discussionmentioning

confidence: 98%

Competition between Serum IgG, IgM, and IgA Anti-Glycan Antibodies

Muthana

Campbell

et al. 2015

PLoS ONE

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Anti-glycan antibodies are an abundant subpopulation of serum antibodies with critical functions in many immune processes. Changes in the levels of these antibodies can occur with the onset of disease, exposure to pathogens, or vaccination. As a result, there has been significant interest in exploiting anti-glycan antibodies as biomarkers for many diseases. Serum contains a mixture of anti-glycan antibodies that can recognize the same antigen, and competition for binding can potentially influence the detection of antibody subpopulations that are more relevant to disease processes. The most abundant antibody isotypes in serum are IgG, IgM, and IgA, but little is known regarding how these different isotypes compete for the same glycan antigen. In this study, we developed a multiplexed glycan microarray assay and applied it to evaluate how different isotypes of anti-glycan antibodies (IgA, IgG, and IgM) compete for printed glycan antigens. While IgG and IgA antibodies typically outcompete IgM for peptide or protein antigens, we found that IgM outcompete IgG and IgA for many glycan antigens. To illustrate the importance of this effect, we provide evidence that IgM competition can account for the unexpected observation that IgG of certain antigen specificities appear to be preferentially transported from mothers to fetuses. We demonstrate that IgM in maternal sera compete with IgG resulting in lower than expected IgG signals. Since cord blood contains very low levels of IgM, competition only affects maternal IgG signals, making it appear as though certain IgG antibodies are higher in cord blood than matched maternal blood. Taken together, the results highlight the importance of competition for studies involving anti-glycan antibodies.

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“…Compared with previous mAbs targeting similar glycans, mAb-A4 appeared to differ in immunohistochemistry staining from RAV12 ( 26 ). Also, anti-HESCA-2 stains primarily a single 250-kDa band on Western blotting of hESC lysate ( 27 ), whereas mAb-A4 stains multiple bands on hESC, indicating that these mAbs target different epitopes (data not shown). FG-88 also did not react with OVCAR3 ( 22 ), whereas mAb-A4 did ( Table 1 ).…”

Section: Discussionmentioning

confidence: 96%

“…A promising category of antigens are the glycans on glycoproteins or glycolipids, which have functional roles in cancer progression ( 7 – 19 ). These glycan antigens can be targeted by monoclonal antibodies (mAb), and indeed, several preclinical anti-glycan mAbs have been developed against OC and other cancers ( 20 – 27 ). However, very few glycan-specific anti-cancer mAbs have had their targets characterized in detail because of difficulties arising from the branched and non-template-driven nature of glycans ( 28 ).…”

Section: Introductionmentioning

confidence: 99%

Characterization of H type 1 and type 1 N-acetyllactosamine glycan epitopes on ovarian cancer specifically recognized by the anti-glycan monoclonal antibody mAb-A4

Choo

Tan

Ding

et al. 2017

Journal of Biological Chemistry

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Cancer-specific glycans of ovarian cancer are promising epitopes for targeting with monoclonal antibodies (mAb). Despite their potential, structural characterization of these glycan epitopes remains a significant challenge in mAb preclinical development. Our group generated the monoclonal antibody mAb-A4 against human embryonic stem cells (hESC), which also bound specifically to N-glycans present on 11 of 19 ovarian cancer (OC) and 8 of 14 breast cancer cell lines tested. Normal cell lines and tissue were unstained by mAb-A4. To characterize the N-linked glycan epitopes on OC cell lines targeted by mAb-A4, we used glycosidases, glycan microarray, siRNA, and advanced high sensitivity matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The mAb-A4 epitopes were found to be Fucα1–2Galβ1–3GlcNAcβ (H type 1) and Galβ1–3GlcNAcβ (type 1 LacNAc). These structures were found to be present on multiple proteins from hESC and OC. Importantly, endo-β-galactosidase coupled with MALDI-MS allowed these two epitopes, for the first time, to be directly identified on the polylactosamines of N-glycans of SKOV3, IGROV1, OV90, and OVCA433. Furthermore, siRNA knockdown of B3GALT5 expression in SKOV3 demonstrated that mAb-A4 binding was dependent on B3GALT5, providing orthogonal evidence of the epitopes' structures. The recognition of oncofetal H type 1 and type 1 LacNAc on OC by mAb-A4 is a novel and promising way to target OC and supports the theory that cancer can acquire stem-like phenotypes. We propose that the orthogonal framework used in this work could be the basis for advancing anti-glycan mAb characterization.

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Anti-Human Embryonic Stem Cell Monoclonal Antibody Hesca-2 Binds to a Glycan Epitope Commonly Found on Carcinomas

Cited by 8 publications

References 53 publications

Therapeutic Antibodies to Ganglioside GD2 Evolved from Highly Selective Germline Antibodies

Therapeutic Antibodies to Ganglioside GD2 Evolved from Highly Selective Germline Antibodies

Competition between Serum IgG, IgM, and IgA Anti-Glycan Antibodies

Characterization of H type 1 and type 1 N-acetyllactosamine glycan epitopes on ovarian cancer specifically recognized by the anti-glycan monoclonal antibody mAb-A4

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