Abstract

In the present study, we have designed a laser-induced fluorescence (LIF) based instrumentation and developed a sensitive methodology for the effective separation, visualization, identification and analysis of proteins on a single platform. In this method, intrinsic fluorescence spectra of proteins were detected after separation on 1 or 2 dimensional Sodium Dodecyl Sulfate–Tris(2-carboxyethyl)phosphine (SDS-TCEP) polyacrylamide gel electrophoresis (PAGE) and the data were analyzed. The MATLAB assisted software was designed for the development of PAGE fingerprint for the visualization of protein after 1- and 2-dimensional protein separation. These provided objective parameters of intrinsic fluorescence intensity, emission peak, molecular weight and isoelectric point using a single platform. Further, the current architecture could differentiate the overlapping proteins in the PAGE gels which otherwise were not identifiable by conventional staining, imaging and tagging methods. Categorization of the proteins based on the presence or absence of tyrosine or tryptophan residues and assigning the corresponding emission peaks (309-356 nm) with pseudo colors allowed the detection of proportion of proteins within the given spectrum. The present methodology doesn't use stains or tags, hence amenable to couple with mass spectroscopic measurements. This technique may have relevance in the field of proteomics that is used for innumerable applications.

Original languageEnglish
Article numbere201700393
JournalJournal of Biophotonics
Volume11
Issue number8
DOIs
Publication statusPublished - 01-08-2018

Fingerprint

Peptide Mapping
interrogation
proteins
Proteins
electrophoresis
Gels
Polyacrylates
Electrophoresis
Polyacrylamide Gel Electrophoresis
gels
phosphine
Fluorescence
Visualization
platforms
methodology
Isoelectric Point
fluorescence
tyrosine
tryptophan
Tryptophan

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Materials Science(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Engineering(all)
  • Physics and Astronomy(all)

Cite this

Siddaramaiah, Manjunath ; Rao, Bola Sadashiva S. ; Joshi, Manjunath B. ; Datta, Anirbit ; Sandya, S. ; Vishnumurthy, Vasudha ; Chandra, Subhash ; Nayak, Subramanya G. ; Satyamoorthy, Kapaettu ; Mahato, Krishna K. / Interrogation of an autofluorescence-based method for protein fingerprinting. In: Journal of Biophotonics. 2018 ; Vol. 11, No. 8.
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abstract = "In the present study, we have designed a laser-induced fluorescence (LIF) based instrumentation and developed a sensitive methodology for the effective separation, visualization, identification and analysis of proteins on a single platform. In this method, intrinsic fluorescence spectra of proteins were detected after separation on 1 or 2 dimensional Sodium Dodecyl Sulfate–Tris(2-carboxyethyl)phosphine (SDS-TCEP) polyacrylamide gel electrophoresis (PAGE) and the data were analyzed. The MATLAB assisted software was designed for the development of PAGE fingerprint for the visualization of protein after 1- and 2-dimensional protein separation. These provided objective parameters of intrinsic fluorescence intensity, emission peak, molecular weight and isoelectric point using a single platform. Further, the current architecture could differentiate the overlapping proteins in the PAGE gels which otherwise were not identifiable by conventional staining, imaging and tagging methods. Categorization of the proteins based on the presence or absence of tyrosine or tryptophan residues and assigning the corresponding emission peaks (309-356 nm) with pseudo colors allowed the detection of proportion of proteins within the given spectrum. The present methodology doesn't use stains or tags, hence amenable to couple with mass spectroscopic measurements. This technique may have relevance in the field of proteomics that is used for innumerable applications.",
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Interrogation of an autofluorescence-based method for protein fingerprinting. / Siddaramaiah, Manjunath; Rao, Bola Sadashiva S.; Joshi, Manjunath B.; Datta, Anirbit; Sandya, S.; Vishnumurthy, Vasudha; Chandra, Subhash; Nayak, Subramanya G.; Satyamoorthy, Kapaettu; Mahato, Krishna K.

In: Journal of Biophotonics, Vol. 11, No. 8, e201700393, 01.08.2018.

Research output: Contribution to journalArticle

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T1 - Interrogation of an autofluorescence-based method for protein fingerprinting

AU - Siddaramaiah, Manjunath

AU - Rao, Bola Sadashiva S.

AU - Joshi, Manjunath B.

AU - Datta, Anirbit

AU - Sandya, S.

AU - Vishnumurthy, Vasudha

AU - Chandra, Subhash

AU - Nayak, Subramanya G.

AU - Satyamoorthy, Kapaettu

AU - Mahato, Krishna K.

PY - 2018/8/1

Y1 - 2018/8/1

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