Antibody variable-region sequencing as a method for hybridoma cell-line authentication.
Cross-contamination and misidentification of various cell lines is a widespread problem that can lead to spurious scientific conclusions. DNA fingerprinting is a powerful identification technique, which can be effectively used for the authentication of human cell lines. In contrast to human cancer cell lines, little attention has so far been given to establishing authentication practices for hybridoma cell lines. Since the majority of hybridomas stem from inbred animals, they have high genetic uniformity, which reduces the applicability of DNA fingerprinting. In the present study, we propose antibody variable-region sequencing as a method of choice for hybridoma cell-line authentication. This method focuses on the most diverse characteristic of hybridoma cell lines and thereby achieves a very high discriminatory power. The sequencing of light-chain variable regions has proven to be especially suitable for routine use because of its high success rate. Two other possible authentication methods, random amplified polymorphic DNA analysis and two-dimensional gel electrophoresis, were also examined. Compared to these and other methods that can be used for discrimination between hybridoma cell lines, variable-region sequencing has many advantages, most notably those of a very high discriminatory power, insensitivity to changes in experimental conditions, simple data analysis, and accessibility to most laboratories.
Koren
Simon
S
Department for Production of Diagnostic Reagents and Research, Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia.
Kosmac
Miha
M
Colja Venturini
Anja
A
Montanic
Sendi
S
Curin Serbec
Vladka
V
D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D006801:Humans; D006825:Hybridomas / Q000166:cytology* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000191:economics / Q000379:methods*
D017931:DNA Primers; D007135:Immunoglobulin Variable Region