Conjugation is determined at pH 12 by measuring PC absorbance at 475nm, using the extinction coefficient of PC= 12.6 x 10E+03. The conjugation ratio is given as a molar ratio of PC to BSA assuming the molecular weight of BSA is 66,430. Store solution in aliquots at -20°C. Protect from light. This product can be dissolved in PBS buffer, water or other neutral buffers at a concentration of 1mg/mL. Product may precipitate when stored in solution, therefore it is recommended to centrifuge the solutions briefly before use and use only the supernatant. When using PBS please refer to the formulation below. PBS Formulation: 8 g NaCl 0.2 g KCl 0.2 g potassium di-hydrogen phosphate (KH2PO4) 1.15 g di-sodium hydrogen phosphate (Na2HPO4) Add to 1 L of water.
Preservative
None
Storage
Store at 2-8°C
Ship
Ambient
Introduction
Phosphorylcholine hapten is conjugated to BSA protein by use of p-diazonium phenylphosphorylcholine.
Citations
Publication ()
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Background
Phosphorylcholine, also known as phosphocholine (ChoP, or PC), is formed by combining a negatively charged phosphate bond with a positively charged choline group, and is expressed in this form by both prokaryotes and eukaryotes. In prokaryotes, ChoP was first identified in the Gram-positive pathogen Streptococcus pneumoniae, and in eukaryotes, ChoP is widespread in fungi, protozoa, insects, and nematodes. ChoP is exposed on the surface of many cells and is often bound to lipids, such as sphingomyelin formed by adding ChoP to ceramides.
Figure 1. Phosphorylcholine moieties found in different biomolecules (Source: Zhang Y, et al. 2023)
ChoP attaches to many surface-exposed bacterial virulence factors. Autolysin, a cell wall degrading enzyme, plays an important role in the growth and cell division of S. pneumoniae. The activity of autolysin is regulated by its interaction with ChoP. In addition, the surface-exposed protein virulence factor PspA of S. pneumoniae is anchored to the pneumococcal surface by a choline interaction with LTA-linked ChoP. Thus, PspA competes with host proteins and inhibits their binding to ChoP, thereby allowing S. pneumoniae to evade complement deposition. In Streptococcus pneumoniae ChoP plays an important role in mediating adhesion and invasion of airway epithelial cells, and it has been found that surface expression of ChoP increases pneumococcal adhesion to human airway cells in vitro and in vivo in mice, thus, ChoP is a common colonization strategy for both Gram-positive and Gram-negative respiratory pathogens.
ChoP plays an important role in regulating host immune responses. In studies of parasite immunomodulation, researchers have found that the ES-62 protein secreted by filarial contains ChoP linked to carbohydrates and has immunomodulatory properties. ES-62 has immunosuppressive effects and may be a potential drug candidate to protect humans against many allergic and autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus and asthma. ES-62 also acts on the production of pathogenic osteoclasts, thereby reducing inflammation and joint damage in a mouse model of collagen-induced arthritis. In addition ChoP has antigenic activity that induces an immune response and prevents respiratory infections.
Alternative Names
phosphocholine PC ChoP
References
1. Zhang Y, et al. The biosynthesis and role of phosphorylcholine in pathogenic and nonpathogenic bacteria. Trends Microbiol. 2023 Jul;31(7):692-706.
2. Frostegård J. Antibodies against phosphorylcholine and protection against atherosclerosis, cardiovascular disease and chronic inflammation. Expert Rev Clin Immunol. 2022 May;18(5):525-532.
Q: Would you mind including a BSA control at the same concentration?
A: BSA control is available, but we only provide it as lyophilized powder (no liquid format).
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References
Longitudinal healing flow diverting stents with phosphorylcholine surface modification
J Neurointerv Surg
Authors: Zoppo CT, Epshtein M, Gounis MJ, Anagnostakou V, King RM.
Background: Flow diversion has become a standard treatment for cerebral aneurysms. However, major drawbacks include the need for dual antiplatelet therapy after implant and delayed complete occlusion of the aneurysm, which occurs when new tissue growth excludes the aneurysm from the parent artery. Biomimetic surface modifications such as the phosphorylcholine polymer (Shield surface modification) represent major advances in reducing thrombogenicity of these devices. However, in vitro studies have raised concerns that this modification may also delay endothelialization of flow diverters.
Methods: Bare metal Pipeline, Pipeline Shield, and Vantage with Shield devices were implanted in the common carotid arteries (CCAs) of 10 rabbits (two in the left CCA, one in the right CCA). Following implant and at 5, 10, 15, and 30 days, the devices were imaged with high-frequency optical coherence tomography and conventional angiography to evaluate tissue growth. At 30 days the devices were explanted and their endothelial growth was assessed with scanning electron microscopy (SEM) at five locations along their length using a semi-quantitative score.
Results: The average tissue growth thickness (ATGT) was not different between the three devices. Neointima was apparent at 5 days and all devices demonstrated similar ATGT at each time point. On SEM, no difference was found in the endothelium scores between the device types.
Conclusion: In vivo, neither the Shield surface modification nor the device design (Vantage) altered the longitudinal healing of the flow diverter.
Circulating choline and phosphocholine measurement by a hydrophilic interaction liquid chromatography-tandem mass spectrometry
Heliyon
Authors: Guerra G, Segrado F, Pasanisi P, Bruno E, Lopez S, Raspagliesi F, Bianchi M, Venturelli E.
Background: Given the growing interest in studying the role of choline and phosphocholine in the development and progression of tumor pathology, in this study we describe the development and validation of a fast and robust method for the simultaneous analysis of choline and phosphocholine in human plasma.
Methods: Choline and phosphocholine quantification in human plasma was obtained using a hydrophilic interaction liquid chromatography-tandem mass spectrometry technique. Assay performance parameters were evaluated using EMA guidelines.
Results: Calibration curve ranged from 0.60 to 38.40 μmol/L (R2 = 0.999) and 0.08-5.43 μmol/L (R2 = 0.998) for choline and phosphocholine, respectively. The Limit Of Detection of the method was 0.06 μmol/L for choline and 0.04 μmol/L for phosphocholine. The coefficient of variation range for intra-assay precision is 2.2-4.1 % (choline) and 3.2-15 % (phosphocholine), and the inter-assay precision range is < 1-6.5 % (choline) and 6.2-20 % (phosphocholine). The accuracy of the method was below the ±20 % benchmarks at all the metabolites concentration levels. In-house plasma pool of apparently healthy adults was tested, and a mean concentration of 15.97 μmol/L for Choline and 0.34 μmol/L for Phosphocholine was quantified.
Conclusions: The developed method shows good reliability in quantifying Choline and Phosphocholine in human plasma for clinical purposes.
COA
Publication 
Figure 1. Phosphorylcholine moieties found in different biomolecules
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