Detection of biotin with zeptomole sensitivity using recombinant spores and a competition assay
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Authors: Lin, Wen-Zhi; Liao, Wan-Chun; Chen, Fang-Hsi; Ma, I-Cheng; Hou, Shao-Yi
Abstract
Detection of protein-binding analytes is important for many applications. Currently, various instrument-based techniques are used for detecting protein-binding analytes. However, such techniques have several limitations including high cost and time-consuming sample processing. In order to overcome these limitations, we developed a sensitive competition assay for the detection of protein-binding analytes using recombinant endospores as a sensing element. The method is based on the competition between the biotin, the model analyte, and a biotin-magnetic bead complex to bind the recombinant spores containing the biotin binding region of streptavidin. After magnetic attraction, the residual spores in the suspension are spread on plates to form colonies which are used to count the amount of the residual spores; the higher the residual ratio of spores, the more biotin in the samples. The linear range was from 150 zmol to 1.5 fmol and the limit of detection of the assay was 150 zmol. The assay proposed herein is sensitive and does not require any expensive equipment. It is suitable for qualitative or semi-quantitative analysis such as screening tests for the detection of toxic chemicals.
Facile Preparation of Phthalocyanine-Based Nanodots for Photoacoustic Imaging and Photothermal Cancer Therapy In Vivo
ACS BIOMATERIALS SCIENCE & ENGINEERING
Authors: Wu, Fengshou; Yue, Liangliang; Cheng, Kai; Chen, Jun; Wong, Ka-Leung; Wong, Wai-Kwok; Zhu, Xunjin
Abstract
The development of near-infrared (NIR)-absorbing nanoagents for personalized multifunctional phototheranostics has attracted considerable attention in the past decade. Recently, the organic nanomaterials with good biosafety are considered as promising phototheranostic agents, while their facile synthesis remains challenging. Inspired by the preparation of carbon nanodots, we fabricate the NIR-absorbing phthalocyanine-based nanodots (ZnPc-NDs) using a facile method for multifunctional phototheranostics. The significant aggregation of phthalocyanines in nanodots induces a complete fluorescence quenching, which affords a high photothermal conversion efficiency (eta = 45.7%). The ZnPc-NDs disperse very well in water media with an average diameter around 80 nm. Further conjugation of biotin on the surface of ZnPc-NDs affords tumor-targeting phthalocyanine nanodots (ZnPc-BT). The ZnPc-BT are demonstrated with favorable biocompatibility, intense photoacoustic signals, high tumor accumulation, and effective tumor suppression in vivo. This Article provides a new insight for further developing nanomedicines with imaging and therapeutic functions to treat cancers precisely and effectively.