Sensitive and selective detection of mycoplasma in cell culture samples using cantilever sensors

✍ Scribed by Sen Xu; Harsh Sharma; Raj Mutharasan

• Publisher: John Wiley and Sons
• Year: 2010
• Tongue: English
• Weight: 807 KB
• Volume: 105
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.22637

✦ Synopsis

Abstract

In this article we report a new biosensor‐based method that is more sensitive and rapid than the current approach for detecting mycoplasma in cell culture samples. Piezoelectric‐excited millimeter‐sized cantilever (PEMC) sensors respond to mass change via resonant frequency change. They are sensitive at femtogram level and can be used directly in liquid for label‐free detection. Common cell culture contaminant, Acholeplasma laidlawii was detected in both buffer and cell culture medium. Two different sources (positive control from a commercial kit and ATCC 23206) were analyzed using antibody‐immobilized PEMC sensor. Resonant frequency decrease caused by binding of A. laidlawii was monitored in real‐time using an impedance analyzer. Positive detection was confirmed by a second antibody binding. The limit of detection (LOD) was lower than 10^3^ CFU/mL in cell culture medium using PEMC sensor while parallel ELISA assays showed LOD as 10^7^ CFU/mL. This study shows that PEMC sensor can be used for sensitive and rapid mycoplasma detection in cell culture samples. Biotechnol. Bioeng. 2010;105: 1069–1077. © 2009 Wiley Periodicals, Inc.