News Release

HemogloBind™ Employed In A Nano LC−MALDI MS/MS Method for Identifying Single Mutation Hemoglobin Variants

MONMOUTH JUNCTION, NJ, February 9, 2023 -- Biotech Support Group reports on a journal article describing the simplicity and efficiency of HemogloBind™, to purify Hemoglobin, in a nano LC−MALDI MS/MS measurement of single amino acid mutations in Hb variants. 

The citation is:

Dasauni, Pushpanjali, et al. "Optimization and Identification of Single Mutation in Hemoglobin Variants with 2, 2, 2 Trifluoroethanol Modified Digestion Method and Nano− LC Coupled MALDI MS/MS." Molecules 27.19 (2022): 6357.

Hemoglobin (Hb) variants arise due to point mutations in globin chains and their pathological treatments rely heavily on the identification of the nature and location of the mutation in the globin chains. Because there is high sequence homology between normal Hb and Hb variant chains, identification of variants by mass spectrometry is very difficult and requires the full sequence coverage of α− and β−chains. Therefore, the present study aims to develop and optimize a specific method of sample processing that could lead to improved sequence coverage and analysis of Hb variants by nano LC−MALDI MS/MS.

The article states “Pure Hb was isolated from hemolysate by using Hemoglobind … The Hb bound to the matrix was eluted by increasing the pH of the buffer using 100 mM Tris borate, pH 9. This allows for easy, one−step isolation of Hb from hemolysate.”

“The use of Hemoglobind reduced the time to obtain pure Hb in an easy single–step procedure. Pure Hb protein is required specifically to optimize and standardize methods for diagnostics. … The desorbed Hb was compatible with LC–MS, and other proteomics studies, as we verified, did not show any change in its intact mass either. This one−step affinity purification gave us the utmost purified Hb.” The article concludes that a sensitive, robust, and reproducible method was developed to identify/ characterize single substitution mutations in the Hb disorder variants. It simply used pure Hb isolated from blood in a single step by using the Hemoglobind matrix, followed by TFE based sample processing and digestion by trypsin, followed by nano−LC separation. The separated peptides were spotted onto MALDI plates upon elution from a nano−LC column and subjected to MALDI MS and MS/MS analysis. The method highlights a non−targeted approach for the identification of Hb variants and its reproducibility to yield 100% sequence coverage for the β−chain with precise identification of all the mutations. It has the potential to become a regular screening/diagnosis tool for the identification of single amino acid mutations in Hb variants. 

“I am very pleased to see that HemogloBind™ performed so well, for such an important heathcare purpose. While most of the applications for HemogloBind™ are for depletion, this application shows its versatility in purifying Hemoglobin for mutational analysis. We envision that this and similar methods will be applicable for not only conventional venous blood samples, but potentially dried blood spots as well, as only small amounts of pure Hemoglobin are required in the final analysis.” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.

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Keywords: Hemoglobin purification, Hemoglobin variants, Hemoglobin mutations, HemogloBind™, Hemoglobinopathies, Sickle cell disease, Thalassemia

About Biotech Support Group LLC

Converging with technological disruptions forthcoming in healthcare, Biotech Support Group continues its growth in the consumable research products area supporting the rapidly expanding installation of LC-MS instrument and computational infrastructure. For this market, key products include: AlbuVoid™ and AlbuSorb™ for albumin depletion, Cleanascite™ for lipid adsorption, HemogloBind™ and HemoVoid™ for hemoglobin removal, For more information, go to http://www.biotechsupportgroup.com.

For Business Development, contact:

Matthew Kuruc 732-274-2866, mkuruc@biotechsupportgroup.com