2016

Journal Applied Surface Science, Volume 362, 30 January 2016, Pages 146153.
Referencia No 17.
CT&F - Ciencia, Tecnologa y Futuro - SYNTHESIS AND ...
www.scielo.org.co/scielo.php?pid=S0122-53832008000100011...
by AY Len-Bermdez - 2008 - Cited by 9 - Related articles
Adan-Yovani Len-Bermdez1* and Ramiro Salazar1. 1Universidad Industrial de Santander
(UIS) - Grupo de Polmeros, Bucaramanga, Santander, Colombia.
https://scholar.google.com.co/scholar
Nueva y por segunda vez citacin bibliogrfica en este ao 2016 por el mismo Juornal Aplplied
Science y por diferentes autores en diferentes artculos (paper), (artculo citado; A Len and R
Salazar CT&F 2008)
en el Journal Applied Surface Science, Volume 362, 30 January 2016, Pages 146153.,
Referencia No 17.,
in Paper; " Microbead-based immunoassay using the outer membrane layer of Escherichia coli
combined with autodisplayed Z-domains".,
By Do-Hoon Kima, Ji-Hong Bonga, Gu Yooa, Seo-Yoon Changa, Min Parka, b, Young Wook
Changa, Min-Jung Kangb, Joachim Josec, Jae-Chul Pyuna,
a Department of Materials Science and Engineering, Yonsei University, 134 Shinchon-dong,
Seodaemun-gu, Seoul 120-749, Republic of Korea
b Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
c Muenster University, Muenster, Germany
Received 6 June 2015, Revised 13 November 2015, Accepted 23 November 2015, Available
online 26 November 2015.
Abstract
The Z-domain has the potential to control the orientation of immobilized antibodies because of
its binding affinity to the Fc regions of antibodies (IgGs). In this work, Z-domains were

autodisplayed on the outer membrane (OM) of Escherichia coli. OM particles were isolated and
coated onto microbeads with positive, neutral, or negative surface charges. Other conditions
such as incubation time and initial OM concentration were also optimized for the OM coating to
obtain maximum antibody-binding. Using three kinds of model proteins with different isoelectric
points (pI), streptavidin (pI = 5, negative charge at pH 7), horseradish peroxidase (pI = 7,
neutral charge at pH 7), and avidin (pI = 10, positive charge at pH 7), protein immobilization
onto the microbeads was carried out through physical adsorption and electrostatic interactions.
Using fluorescently labeled antibodies and fluorescence-activated cell sorting, it was
determined that the neutral and the positively charged microbeads effectively bound antibodies
while minimizing non-specific protein binding. The OM-coated microbeads with autodisplayed
Z-domains were applied to C-reactive protein immunoassay. This immunoassay achieved 5fold improved sensitivity compared to conventional immunoassay based on physical adsorption
of antibodies at the cutoff concentration of medical diagnosis of inflammatory diseases (1000
ng/ml) and cardiovascular diseases (200 ng/ml).
http://www.sciencedirect.com//article/pii/S0169433215029281
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