Uzma Naseeb1, Jonas Axelsson2, Theres Jägerbrink3, Jawed Shafqat3, Shamshad Zarina4, Hans Jörnvall3. 1. Department of Medical Biochemistry and Biophysics / Clinical Science, Intervention and Technology2, Karolinska Institutet, Stockholm, Sweden. / National Center for Proteomics, University of Karachi, Karachi. 2. Department of Medical Biochemistry and Biophysics / Clinical Science, Intervention and Technology2, Karolinska Institutet, Stockholm, Sweden. 3. Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden. 4. National Center for Proteomics, University of Karachi, Karachi.
Abstract
OBJECTIVE: To complement an earlier analysis of protein alterations in plasma from uremic versus healthy subjects by addition of further LC-MS/MS analysis to the previously used MALDI-TOF mass analyses. METHODOLOGY: Sequence identifications of tryptic peptides from SDS gel electrophoretic fractions of immunodepleted and HPLC-fractionated plasma was performed from seven chronic kidney disease stage 5 patients (age 55 ± 14 years, glomerular filtration rate 6.9 ±2.9 mL/minute/1.73 m2) and from seven matched controls. RESULTS: About twice as many proteins were increased in uremic plasma as the previously identified. The identifications included proteins that consistently complement the two identification patterns regarding separate subunits from the same protein complex. CONCLUSION: Mass spectrometric analysis is applicable to complex plasma proteomes in clinical settings. The LC-MS/MS technique, based on individual peptide sequence analyses, gives increased identifications and also demonstrates feasibility of this technique in clinical practice.
OBJECTIVE: To complement an earlier analysis of protein alterations in plasma from uremic versus healthy subjects by addition of further LC-MS/MS analysis to the previously used MALDI-TOF mass analyses. METHODOLOGY: Sequence identifications of tryptic peptides from SDS gel electrophoretic fractions of immunodepleted and HPLC-fractionated plasma was performed from seven chronic kidney disease stage 5 patients (age 55 ± 14 years, glomerular filtration rate 6.9 ±2.9 mL/minute/1.73 m2) and from seven matched controls. RESULTS: About twice as many proteins were increased in uremic plasma as the previously identified. The identifications included proteins that consistently complement the two identification patterns regarding separate subunits from the same protein complex. CONCLUSION: Mass spectrometric analysis is applicable to complex plasma proteomes in clinical settings. The LC-MS/MS technique, based on individual peptide sequence analyses, gives increased identifications and also demonstrates feasibility of this technique in clinical practice.
Authors: Jack Gisby; Candice L Clarke; Nicholas Medjeral-Thomas; Michelle Willicombe; David C Thomas; James E Peters; Talat H Malik; Artemis Papadaki; Paige M Mortimer; Norzawani B Buang; Shanice Lewis; Marie Pereira; Frederic Toulza; Ester Fagnano; Marie-Anne Mawhin; Emma E Dutton; Lunnathaya Tapeng; Arianne C Richard; Paul Dw Kirk; Jacques Behmoaras; Eleanor Sandhu; Stephen P McAdoo; Maria F Prendecki; Matthew C Pickering; Marina Botto Journal: Elife Date: 2021-03-11 Impact factor: 8.713