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Biotech Support Group Release: Synovial Fluid Biomarker Discovery Sample Preparation Utilizing Albusorb™ for Albumin Depletion



9/4/2012 10:55:15 AM

PRLog (Press Release) - Sep 02, 2012 - MONMOUTH JUNCTION, NJ.– Approximately 65% of the total protein in normal synovial fluid consists of human serum albumin. Authors Kaisa E. Happonen et al published an article titled, “PRELP inhibits the formation of the complement membrane attack complex” in the Journal of Biological Chemistry which cites AlbuSorb™ from Biotech Support Group for albumin depletion from synovial fluid for detecting and measuring Proline arginine rich end leucine-rich repeat protein (PRELP). Scientists demonstrated that PRELP inhibits MAC by decreasing C9 polymerization, thereby preventing limiting complement attack on basement membranes. Proper identification of low-abundance proteins in synovial fluid that may prevent disease biomarkers discovery is enhanced by albumin depletion. After albumin depletion, synovial fluid samples are concentrated using trhichloroacetic acid and incubated. Dried precipitates are separated after pelleting, washing, reducing on SDS-PAGE buffer and boiling on gels. Proteins on polyvinylidene fluoride membranes are incubated with a 125I-labeled polyclonal anti-PRELP antibody and antibodies bound are compared to pre-incubated antibody with PRELP. Thus, joint disease cartilage destruction results in PRELP proliferation into synovial fluid which is measured using ELISA.

Proteomic biotechniques allow for the analysis of proteins in synovial fluid. After using Albusorb™, synovial fluid proteins on two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) is studied. Abundant proteins HSA and IgG are depleted using Albusorb™, followed by size-exclusion chromatography (SEC) of intact proteins prior to preparation of samples for analysis by LC/LC-MS/MS. Next tandem mass spectrometry (MS/MS), coupled with multidimensional liquid chromatography (LC) and database searching is effective technique for protein identification and characterization. Examples of elevated biomarker candidates which are elevated in erosive or nonerosive rheumatoid arthritis (RA) are G3PDH,Peptidylprolyl isomerase, Cystatin B, Phosphoglycerate mutase 1, a2-plasmin inhibitor, S100A8 (calgranulin A), IgG1H Nie, Thymosin-ß4. The ultimate goal of such application would be to comparatively find and analyze novel biomarkers of rheumatoid arthritis and search for the same biomarkers in synovial fluid of disease and non disease patient samples by proteomic techniques.

Characteristics of Albusorb™

Removes >90% albumin from 30 mg albumin/ml sample

Affinity-type equivalence, virtually no cross-reactivity with other proteins

Bind and elute procedure - simply weigh powder, condition the sample, centrifuge and/or filter, and recover the albumin depleted serum

Economical new surface technology, not based on affinity chromatography

Mild conditions maintain tertiary structure of proteins and simple transfer to secondary analysis

The albumin depleted filtrate retains the enzymatic and biological activity

Removes albumin from samples such serum, plasma and from species including human, mouse, sheep, bovine, goat, rat, and calf

The flow through fraction is compatible with LC-MS, activity based protein profiling and proteomic studies.

Biotech Support Group is interested in collaborating on research and development of peptide inhibitors of the complement system by using Albusorb™ for sample preparation.

Biotech Support Group also has AlbuVoid™ for albumin depletion plus low abundance serum protein enrichment.

For more information, visit:

AlbuSorb™ Albumin Depletion Kit

http://www.biotechsupportgroup.com/node/64

AlbuVoid™ - Albumin Depletion and Low Abundance Protein Enrichment Kit from Serum or Plasma

http://www.biotechsupportgroup.com/node/55

About Biotech Support Group LLC

Biotech Support Group LLC is a leading provider of genomics and proteomics sample preparation products and enrichment reagent kits as well as integrated biotechnology services for life sciences research, biomarker and drug discovery. Based in New Jersey, it’s principal products include: AlbuVoid™ for albumin depletion, Cleanascite™ for lipid adsorption and clarification, NuGel™ for passivated silica-based affinity chromatography, and ProCipitate™ & ProPrep™ for nucleic acid isolation. Currently, Biotech Support Group LLC and ProFACT Proteomics Inc., are collaborating on the development of a proteomics platform used in functional profiling for proteomic analysis and a separations method for generating sub-proteomes used in biomarker and functional proteomic prospecting. For more information, go to: www.biotechsupportgroup.com

CONTACT:

Dr.Swapan Roy

Biotech Support Group

1 Deer Park Drive, Suite M,

Monmouth Junction, NJ 08852, USA

732-274-2866

sales@biotechsupportgroup.com

Albusorb™ References:

Gwenael Pottiez, Pawel Ciborowski. Proteomic Profiling of Cerebrospinal Fluid Expression Profiling In Neuroscience Neuromethods.2012;64:245-270

Happonen KE, Fürst CM, Saxne T et al. PRELP protein inhibits the formation of the complement membrane attack complex.Journal of Biological Chemistry.2012;287(11):8092-100

Holmberg R, Refai E, Höög A.Lowering apolipoprotein CIII delays onset of type 1 diabetes. Proceedings of the National Academy of Sciences.2011;108(26):10685-9.

Tang MX, Ogawa K, Asamoto M. Effects of Nobiletin on PhIP-Induced Prostate and Colon Carcinogenesis in F344 Rats Nutrition and Cancer.2011;63(2):227-33

Holmberg, Rebecka Apolipoprotein CIII and Ljungan virus in diabetes 2010. Doctoral Thesis

Lu Q, Zheng X, McIntosh T Development of different analysis platforms with LC-MS for pharmacokinetic studies of protein drugs. Analytical Chemistry.2009;81(21):8715-23

References

Nandakumar, K. S., Jansson, A., Xu, B., Rydell, N., Ahooghalandari, P., Hellman, L., Blom, A. M., and Holmdahl, R. PLoS One.2010; 5(10):e13511

Blom, A. M., Nandakumar, K. S., and Holmdahl, R. Annals of the Rheumatic Diseases.2009;68(1):136-142

Gibson, D. S. and Rooney, M. E. The human synovial fluid proteome: A key factor in the pathology of joint disease. Proteomics Clinical Applications.2007; 1: 889–899

Hu, S., Loo, J. A. and Wong, D. T. Human body fluid proteome analysis. Proteomics .2006;6:6326-6353

Liao, H., Wu, J., Kuhn, E., Chin, W., Chang, B., Jones, M. D., O'Neil, S., Clauser, K. R., Karl, J., Hasler, F., Roubenoff, R., Zolg, W. and Guild, B. C. Use of mass spectrometry to identify protein biomarkers of disease severity in the synovial fluid and serum of patients with rheumatoid arthritis.2004:Arthritis & Rheumatism; 50:3792-3803

Jikimoto T, Nishikubo Y, Koshiba M, Kanagawa S, Morinobu S, Morinobu A, et al. Thioredoxin as a biomarker for oxidative stress in patients with rheumatoid arthritis. Molecular Immunology. 2002;38: 765-72

Wu SL, Amato H, Biringer R, Choudhary G, Shieh P, Hancock WS. Targeted proteomics of low-level proteins in human

plasma by LC/MSn: using human growth hormone as a model system. Journal of Proteomic Research; 2002;1:459-65. Sinz A, Bantscheff M, Mikkat S, Ringel B, Drynda S, Kekow J, et al. Mass spectrometric proteome analyses of synovial fluids and plasmas from patients suffering from rheumatoid arthritis and comparison to reactive arthritis or osteoarthritis. Electrophoresis.2002; 23: 3445-56.

Uchida T, Fukawa A, Uchida M, Fujita K, Saito K. Application of a novel protein biochip technology for detection and identification of rheumatoid arthritis biomarkers in synovial fluid. Journal of Proteomic Research.2002; 1: 495–9

Wang, Y., Rollins, S. A., Madri, J. A., and Matis, L. A.Proceedings of the National Academy of Sciences of the United States of America.1995;92(19):8955-8959

Weinberger A, Simkin PA. Plasma proteins in synovial fluids of normal human joints. Seminars in Arthritis and Rheumatism 1989; 19:66-76


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