Kinetics of Antibody Binding to Membranes of Living Bacteria Measured by a Photonic Crystal-Based Biosensor
BIOSENSORS-BASEL
Authors: Rostova, Ekaterina; Ben Adiba, Carine; Dietler, Giovanni; Sekatskii, Sergey K.
Abstract
Optical biosensors based on photonic crystal surface waves (PC SWs) offer a possibility to study binding interactions with living cells, overcoming the limitation of rather small evanescent field penetration depth into a sample medium that is characteristic for typical optical biosensors. Besides this, simultaneous excitation of s-and p-polarized surface waves with different penetration depths is realized here, permitting unambiguous separation of surface and volume contributions to the measured signal. PC-based biosensors do not require a bulk signal correction, compared to widely used surface plasmon resonance-based devices. We developed a chitosan-based protocol of PC chip functionalization for bacterial attachment and performed experiments on antibody binding to living bacteria measured in real time by the PCSW-based biosensor. Data analysis reveals specific binding and gives the value of the dissociation constant for monoclonal antibodies (IgG2b) against bacterial lipopolysaccharides equal to K-D = 6.2 +/- 3.4 nM. To our knowledge, this is a first demonstration of antibody-binding kinetics to living bacteria by a label-free optical biosensor.
Immunoadjuvant activity in mice of polysaccharides isolated from the leaves of Panax ginseng CA Meyer
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Authors: Hwang, Su-Hyun; Shin, Myoung-Sook; Yoon, Taek Joon; Shin, Kwang-Soon
Abstract
Our previous study showed polysaccharide (GS-P) isolated from the leaves of Panax ginseng C.A. Meyer possessed anti-tumor metastatic activity in mouse model. In this study, we evaluated the immunoadjuvant effect of GS-P on the induction of humoral and cellular immune responses against ovalbumin (OVA) in mice. When mice were immunized subcutaneously with OVA admixed with or without GS-P, the OVA+GS-P group showed significantly higher antibody production than the group immunized with OVA alone. This suggests that GS-P has the ability to enhance the adaptive immune response. In addition, the OVA+GS-P+FIA (Freund's incomplete adjuvant) group induced higher levels of antigen-specific IgG1 and IgG2b antibodies than the OVA+FIA group. The culture supernatant obtained from the splenocytes of mice immunized with OVA+GS-P+FIA showed higher levels of OVA-specific Thl-type (IL-2, IFN-gamma, GM-CSF) and Th2-type (IL-10) cytokines. Following in vitro analysis of T cell proliferation, the splenocytes of mice treated with OVA+GS-P+FIA showed significantly more proliferation than those treated with OVA+FIA. Further, the production of IgE antibody was dramatically reduced when OVA+GS-P+FIA was used to immunize mice rather than OVA+FIA or OVA+FCA (Freund's complete adjuvant). Collectively, these results suggest that GS-P may possess adjuvant activity that potentially enhances humoral as well as cellular immune responses. (C) 2017 Elsevier B.V. All rights reserved.