Paraffin-embedded cell line microarray (PECLIMA): development and validation of a high-throughput method for antigen profiling of cell lines.
The introduction of high-throughput techniques is increasingly providing abundant information on molecular alterations requiring validation at the posttranscriptional level. Protein expression is now efficiently evaluated in large series of tumors included in tissue microarrays. We propose, describe and validate a technique to elaborate paraffin-embedded cell line microarrays (PECLIMA) from fixed cell cultures, which can be processed like standard surgical pathology biopsies prior to immunophenotyping. Our results show a reliable protein immunoexpression profiling in six widely used cell lines under different fixation conditions. This technique permits the simultaneous analysis of multiple antigens in multiple cell lines under different experimental conditions. Additional features of these arrays are long-term storage, their suitability for a variety of techniques including immunocytochemistry and in situ hybridization and their low cost.
Ferrer
Berta
B
Department of Anatomical Pathology, Institut d'Investigacions Biomèdiques August Pi i Sunyer, Hospital Clinic and University of Barcelona, Barcelona, Spain.
Bermudo
Raquel
R
Thomson
Timothy
T
Nayach
Iracema
I
Soler
Marta
M
Sánchez
Montserrat
M
Castillo
Mireia
M
Calvo
Julia
J
Campo
Elias
E
Fernández
Pedro L
PL
D000951:Antigens, Neoplasm / Q000032:analysis* / Q000276:immunology; D014408:Biomarkers, Tumor / Q000378:metabolism; D045744:Cell Line, Tumor / Q000378:metabolism*; D016591:Histocytological Preparation Techniques / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D016612:Paraffin Embedding; D010641:Phenotype; D040081:Protein Array Analysis / Q000379:methods*
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor