TY - JOUR
T1 - Bifunctional nickel-iminodiacetic acid-core-shell silica nanoparticles for the exclusion of high molecular weight proteins and purification of His-tagged recombinant proteins
AU - Hernandez-Leon, Sergio G.
AU - Sarabia-Sainz, Jose A.
AU - Ramos-Clamont Montfort, Gabriela
AU - Huerta-Ocampo, José Ángel
AU - Guzman-Partida, Ana M.
AU - Robles-Burgueño, Maria del Refugio
AU - Burgara-Estrella, Alexel J.
AU - Vazquez-Moreno, Luz
N1 - Publisher Copyright:
© The Royal Society of Chemistry.
PY - 2019
Y1 - 2019
N2 - Herein, silica nanoparticles were synthesized and chemically modified with iminodiacetic acid (IDA) and Ni2+ ions surrounded by a bis-acrylamide polymeric shell to obtain a new core-shell immobilized metal affinity chromatography (IMAC) based material. These Ni2+-IDA-core-shell silica nanoparticles (Ni2+-IDA-CSS-NP) represent a new alternative for purification of His-tagged proteins and exclusion of high molecular weight (HMW) proteins at the same time. Nanoparticles presented a final size of 479.6 ± 6.9 nm determined by dynamic light scattering (DLS) and a surface charge of −37.2 ± 0.5 mV. Successful incorporation of the different compounds at every phase of synthesis was evidenced by ATR-FTIR analysis. Ni2+-IDA-CSS-NP were used for isolation of His-tagged spo0F (6His-spo0F) from E. coli lysate. Ni2+-IDA-CSS-NP presented a capacity of 4.16 ± 0.45 μg mg−1. Purification of 6His-spo0F with high selectivity and the effective exclusion of HMW proteins were evidenced by SDS-PAGE and validated through mass spectrometry analysis.
AB - Herein, silica nanoparticles were synthesized and chemically modified with iminodiacetic acid (IDA) and Ni2+ ions surrounded by a bis-acrylamide polymeric shell to obtain a new core-shell immobilized metal affinity chromatography (IMAC) based material. These Ni2+-IDA-core-shell silica nanoparticles (Ni2+-IDA-CSS-NP) represent a new alternative for purification of His-tagged proteins and exclusion of high molecular weight (HMW) proteins at the same time. Nanoparticles presented a final size of 479.6 ± 6.9 nm determined by dynamic light scattering (DLS) and a surface charge of −37.2 ± 0.5 mV. Successful incorporation of the different compounds at every phase of synthesis was evidenced by ATR-FTIR analysis. Ni2+-IDA-CSS-NP were used for isolation of His-tagged spo0F (6His-spo0F) from E. coli lysate. Ni2+-IDA-CSS-NP presented a capacity of 4.16 ± 0.45 μg mg−1. Purification of 6His-spo0F with high selectivity and the effective exclusion of HMW proteins were evidenced by SDS-PAGE and validated through mass spectrometry analysis.
UR - http://www.scopus.com/inward/record.url?scp=85064246871&partnerID=8YFLogxK
U2 - 10.1039/C9RA01144G
DO - 10.1039/C9RA01144G
M3 - Artículo
AN - SCOPUS:85064246871
SN - 2046-2069
VL - 9
SP - 11038
EP - 11045
JO - RSC Advances
JF - RSC Advances
IS - 20
ER -