GYPA Recombinant Monoclonal Antibody
Description
Definition and Biological Significance
GYPA (glycophorin A) is a transmembrane protein encoded by the GYPA gene (UniProt ID: P02724) that determines M/N blood group antigens and contributes to erythrocyte mechanical stability . The recombinant monoclonal antibody against GYPA is engineered to bind specifically to this protein, facilitating studies on red blood cell biology, blood group typing, and pathogen-host interactions .
Production Process
The antibody is synthesized via a recombinant pipeline:
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Gene Cloning: GYPA-specific antibody genes are isolated and inserted into mammalian expression vectors .
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Host Cell Expression: Genes are transfected into cells (e.g., HEK293) for antibody synthesis .
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Purification: Affinity chromatography (e.g., Protein A) ensures high purity .
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Validation: ELISA, Western blot, and flow cytometry confirm specificity and affinity .
This method eliminates hybridoma variability and enables scalable production .
Key Applications
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Erythrocyte Marker: Detects red blood cells and erythroid precursors in IHC and flow cytometry .
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Blood Group Typing: Identifies M/N antigens for transfusion compatibility studies .
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Infectious Disease Research: Serves as a receptor for pathogens like Sandei virus and Streptococcus adhesins .
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Multiplex Assays: Conjugation-ready formats enable simultaneous detection in cytometric bead arrays .
Performance Data
Advances in Engineering
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Subclass Switching: IgG subclass variants (e.g., IgG1, IgG2a) enable multiplex labeling using subclass-specific secondary antibodies .
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Glycosylation Optimization: Bisecting GlcNAc modifications enhance antibody-dependent cellular cytotoxicity (ADCC) by 3-fold .
Production Challenges
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Post-Translational Modifications: Citric acid in formulation buffers may cause covalent modifications (e.g., +174 Da adducts), increasing heterogeneity .
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Batch Consistency: Requires stringent control of glycosylation patterns to maintain functional stability .
Future Directions
Ongoing research focuses on:
Product Specs
The GYPA recombinant monoclonal antibody is produced through a sophisticated process involving the acquisition of GYPA antibody genes. These genes are then introduced into carefully selected host cells, which are cultured to synthesize the antibodies using a cell-based expression and translation system. This approach offers notable advantages, including enhanced purity and stability of the resulting GYPA recombinant monoclonal antibodies, while also improving their affinity and specificity.
Following synthesis, the GYPA recombinant monoclonal antibody undergoes a rigorous purification process utilizing affinity chromatography. Subsequently, it undergoes comprehensive testing through ELISA and FC assays. This antibody is highly specific, targeting only the human GYPA protein.
GYPA (glycophorin A) plays a critical role as a key component of the erythrocyte membrane, contributing to the maintenance of membrane structure, blood group determination, mechanical resilience, and potentially cell adhesion and signaling. Its presence on the surface of red blood cells is essential for their proper function in oxygen transport and maintaining blood compatibility.
Target Background
- Studies have demonstrated differential sensitivities of surface proteins on K562 cells to proteases, suggesting molecular mechanisms of transmembrane protein transport and cycling. PMID: 28980921
- A complex structural rearrangement involving the loss of GYPB and the acquisition of two GYPB-A hybrid genes has been linked to a 40% reduction in severe malaria risk and has recently increased in frequency in certain regions of Kenya. PMID: 28522690
- Research has shown that Plasmodium falciparum EBA-175 binding to glycophorin A triggers a phosphorylation cascade involving erythrocyte cytoskeletal proteins, leading to changes in the viscoelastic properties of the host cell. PMID: 28226242
- While weak interactions between glycophorin and band 3 are undeniable, glycophorin A and band 3 appear to have separate interactions in the membrane that regulate their lateral mobility. PMID: 27580023
- CD235a has emerged as a potential highly sensitive predictor, similar to CD4. PMID: 25716234
- Glycophorin-A-rich microparticles released from evolving growing thrombi into the distal perfusing blood can be detected in peripheral blood. CD235a(+) cMPs may represent a novel systemic biomarker of ongoing thrombosis. PMID: 26239059
- Studies have demonstrated that the initial vacuolar membrane surrounding internalized Babesia divergens is formed from protein and lipid components of the red blood cell plasma membrane, including band 3, glycophorin A, and spectrin. PMID: 25628009
- Erythrocytes deficient in GPA and band 3 exhibit resistance to malaria infection. PMID: 25778531
- A genotype-serotype correlation study in Taiwan has classified ten alleles of the MN blood group system, highlighting major and minor variations of the standard alleles. PMID: 24858913
- PfEBA-175 engages multiple glycans of GpA encoded by exon 3, suggesting that the presentation of glycans is crucial for high-avidity binding. PMID: 25205096
- Local increases in the expression of glycophorin A and osteoprotegerin have been observed in carotid atherosclerotic lesions of symptomatic patients compared to asymptomatic individuals. PMID: 23722820
- Research has highlighted the importance of regions within PfEBA175, beyond the DBL domains, in the interaction with GYPA, emphasizing their potential inclusion in an EBA175-based vaccine. PMID: 24043627
- GPA-NN mutagens have been implicated in the pathogenesis of chronic benzene poisoning. PMID: 17456399
- Beta-branched residues adjacent to GG4 motifs contribute to the efficient association of glycophorin A transmembrane helices. PMID: 21072853
- Data suggest that glycophorin A (Gpa) enhances the expression and activity of Cl-/HCO3- exchanger Ae1, while a G719D mutation renders Ae1 mutant constructs unresponsive to GPA. This suggests a role for Ae1 amino acids 22-28 in GPA responsiveness. PMID: 21455273
- The GG genotype of the rs 1489759 HHIP single-nucleotide polymorphism (SNP) and the CC genotype of the rs 2202507 GYPA SNP have been associated with a protective effect against COPD and lung cancer. PMID: 21119205
- Studies indicate that the lipid bilayer thickness significantly influences the monomer-dimer equilibrium of the glycophorin A transmembrane domain, potentially altering its dimerization propensities. PMID: 20603102
- ABH blood group antigens have been identified in the N-glycan of human glycophorin A. PMID: 20434428
- A Sta gene in a specific family has been characterized with A246 & C252, C302, and G307. The putative crossing-over point is located between nucleotides 252 to 302 in Intron 3. PMID: 20233359
- Energy transfer between differently labeled glycophorin A transmembrane helices has been observed to decrease with increasing Sodium Dodecyl Sulfate mole fractions without altering the helicity of the peptides. PMID: 20074546
- A heterozygous missense mutation E758K in the human AE1/SLC4A1/band 3 gene has been identified in two unrelated patients with well-compensated hereditary spherostomatocytic anemia. This mutation induces glycophorin A-independent, endogenous cation transport. PMID: 19907019
- Research has highlighted the role of natural selection in shaping polymorphisms/variants on the erythrocyte surface. PMID: 11861881
- Somatic cell mutations at the gene locus in erythrocytes of radiation workers at a nuclear facility have been reported. PMID: 12492375
- Distinct regions of human glycophorin A have been shown to enhance human red cell anion exchanger transport function and surface trafficking. PMID: 12813056
- Band 3 in red blood cells can adopt two distinct structures: one with high anion transport activity when glycophorin A is present and another with lower activity when glycophorin A is absent. PMID: 14604989
- Porcine Kupffer cells bind xenogeneic human RBC through recognition of a carbohydrate epitope on glycophorin A. PMID: 16003235
- This study provides a systematic classification of ten alleles (five M and five N alleles) of the MN blood group system, categorizing them into major or minor variations of the standard alleles. PMID: 16205834
- Somatic mutation frequency levels of glycophorin A in newborns have been found not to be related to maternal lifestyle factors or drug metabolizing enzyme genotypes. PMID: 16424825
- An analysis of GYPA & GYPB genotypes carrying MNSs antigens in Thai patients with cerebral malaria and mild malaria did not reveal any statistical difference, suggesting that MNSs antigens might not be significant in susceptibility to cerebral malaria. PMID: 17372674
- Single mutations in glycophorin A have been shown to result in the observed fourth power relationship with age of oncet in polycythemia rubra vera. PMID: 17452518
- GPA depletion does not alter the native conformation of band 3 at the DIDS binding site. However, it modulates a conformational equilibrium between two states of the binary complex formed by the competitive inhibitor DIDS, reversibly bound to properly folded band 3. PMID: 19071041
- AE1 and GPA have been found to form a complex in the endoplasmic reticulum of human K562 cells. PMID: 19438409
