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Quantifying proteins at microgram levels integrating gel electrophoresis and smartphone technology

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Autor(es):
de Jesus, Jemmyson Romario [1, 2] ; Guimaraes, Ivanilce Cristina [1, 2] ; Zezzi Arruda, Marco Aurelio [1, 2]
Número total de Autores: 3
Afiliação do(s) autor(es):
[1] Natl Inst Sci & Technol Bioanalyt, Campinas, SP - Brazil
[2] Univ Campinas UNICAMP, Spectrometry Sample Preparat & Mechanizat Grp GEP, Inst Chem, Campinas, SP - Brazil
Número total de Afiliações: 2
Tipo de documento: Artigo Científico
Fonte: JOURNAL OF PROTEOMICS; v. 198, p. 45-49, APR 30 2019.
Citações Web of Science: 0
Resumo

A combination between gel electrophoresis and smartphone technology is applied for quantifying proteins in a serum sample. Electrophoresis not only allows one to separate different proteins but also to build-up a calibration curve for those proteins. To expand its applicability, a smartphone allows one to capture a gel image, through its camera. Additionally, the treatment of the data extracted through each protein band is performed using a freely available program (PhotoMetrix), which is downloaded to one's own smartphone. Through this approach, the quantification of proteins such as albumin, immunoglobulin, and carbonic anhydrase is performed in a serum sample by acquiring images using 32 x 32 pixels for each image in the region of the protein bands. An LOQ from 1.9 to 2.4 mu g and r(2) > 0.980 may be obtained. Comparing results through the analyses of the gels using Image Master 2D Platinum 6.0 software reflects good agreement (95% confidence level) between the results. Significance statement In many biological studies involving structural analysis or biophysical and biochemical characterization after purification process, or even in assays of protein:protein interaction, the estimation of protein quantity and protein purity is a fundamental step. However, even although many methodologies are proposed in the literature for quantifying proteins, they present some limitations, once are frequently expensive and require solution sample. In addition, they usually do not quantify the specific active concentration, but the total amount. Thus, it is necessary to develop an easy, fast and low-cost method that allows to quantify and to evaluate protein purity of reactional system, for example, a protein:protein interaction. In this way, we presented a simple strategy based on the integration of Smartphone technology and gel electrophoresis, where SDS-PAGE provides multiple information regarding the quality of the protein, such as the presence of degradation products, as well as if there was interaction reaction between proteins. Then, the smartphone detects the proteins in the SDS-PAGE gel, allowing to evaluate purity degree and the quantity, at microgram level, of the protein. We believe that the combination of these features may help to increase the productivity and accuracy of gel electrophoresis analysis, and the results obtained through a smartphone, easily achieved in our pockets. (AU)

Processo FAPESP: 16/07384-7 - Bioimageamento elementar por meio da ablação a laser e espectrometria de massas com plasma indutivamente acoplado
Beneficiário:Marco Aurelio Zezzi Arruda
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 18/00768-0 - Correlação entre metais e homeostase proteica: estudos iniciais investigando a incorporação do isótopo 68Zn na co-chaperona Hsp40 e seu efeito em levedura
Beneficiário:Jemmyson Romário de Jesus
Linha de fomento: Bolsas no Brasil - Pós-Doutorado