Comparative analysis of electrophoresis and interferometric optical detection method for molecular weight determination of proteins
B. Santamaría,
A. L. Hernandez,
M.F. Laguna,
M. Holgado
Affiliations
B. Santamaría
Group of Optics, Photonics, and Biophotonics, Center for Biomedical Technology (CTB), Universidad Politécnica de Madrid, Parque Científico y Tecnológico de la UPM, Campus de Montegancedo, M-40 km38, 28223 Pozuelo de Alarcón, Madrid, Spain; Group of Organ and Tissue on-a-chip and In-Vitro Detection, Health Research Institute of the Hospital Clínico San Carlos, IdISSC. C/ Profesor Martín Lagos s/n, 4a Planta Sur 28040, Madrid, Spain; Department of Mechanics, Chemistry and Industrial Design Engineering, Escuela Superior de Ingeniería y Diseño Industrial, Universidad Politécnica de Madrid, Ronda de Valencia 3, 28012, Madrid, Spain
A. L. Hernandez
Group of Optics, Photonics, and Biophotonics, Center for Biomedical Technology (CTB), Universidad Politécnica de Madrid, Parque Científico y Tecnológico de la UPM, Campus de Montegancedo, M-40 km38, 28223 Pozuelo de Alarcón, Madrid, Spain; Group of Organ and Tissue on-a-chip and In-Vitro Detection, Health Research Institute of the Hospital Clínico San Carlos, IdISSC. C/ Profesor Martín Lagos s/n, 4a Planta Sur 28040, Madrid, Spain
M.F. Laguna
Group of Optics, Photonics, and Biophotonics, Center for Biomedical Technology (CTB), Universidad Politécnica de Madrid, Parque Científico y Tecnológico de la UPM, Campus de Montegancedo, M-40 km38, 28223 Pozuelo de Alarcón, Madrid, Spain; Group of Organ and Tissue on-a-chip and In-Vitro Detection, Health Research Institute of the Hospital Clínico San Carlos, IdISSC. C/ Profesor Martín Lagos s/n, 4a Planta Sur 28040, Madrid, Spain; Department of Applied Physics and Materials Engineering, Escuela Técnica Superior de Ingenieros Industriales, Universidad Politécnica de Madrid, C/José Gutiérrez Abascal, 2, 28006, Madrid, Spain
M. Holgado
Group of Optics, Photonics, and Biophotonics, Center for Biomedical Technology (CTB), Universidad Politécnica de Madrid, Parque Científico y Tecnológico de la UPM, Campus de Montegancedo, M-40 km38, 28223 Pozuelo de Alarcón, Madrid, Spain; Group of Organ and Tissue on-a-chip and In-Vitro Detection, Health Research Institute of the Hospital Clínico San Carlos, IdISSC. C/ Profesor Martín Lagos s/n, 4a Planta Sur 28040, Madrid, Spain; Department of Applied Physics and Materials Engineering, Escuela Técnica Superior de Ingenieros Industriales, Universidad Politécnica de Madrid, C/José Gutiérrez Abascal, 2, 28006, Madrid, Spain; Corresponding author. Group of Optics, Photonics, and Biophotonics, Center for Biomedical Technology (CTB), Universidad Politécnica de Madrid, Parque Científico y Tecnológico de la UPM, Campus de Montegancedo, M-40 km38, 28223 Pozuelo de Alarcón, Madrid, Spain.
Analytical detection methods play a pivotal role in scientific research, enabling the identification and quantification of specific analytes in various disciplines. This scientific report aims to compare two very different methodologies for determining the Molecular Mass (MM, also known as Molecular Weight, MW) of proteins: electrophoresis gel and the Interferometric Optical Detection Method (IODM). For this purpose, several proteins with different MM were selected.The electrophoresis technique was employed to validate the structure and MM of different parts or fragments of the Matrix Metallopeptidase 9 antibody (anti-MMP9), antibody against S100 calcium binding protein A6 (anti-S100A6) and Cystatin S4 antibody (anti-CST4) by examining the presence of bands with expected sizes. The IODM was applied to study the above-mentioned proteins (part of the antibodies) together with the protein G, as a reference to correlate the MM and protein sizes with the measured signal.We report the evidence of IODM as a competitive analytical approach for the determination of the MM of proteins for the first time. This innovative method allows for accurate MM determination using minimal sample volumes and concentrations, employing a simple experimental procedure that eliminates the requirement for protein denaturation.
