Microsampling for RNA biomarker profiling in blood
Poster number: 165
Presenter: Les Kobzik, Ph.D., Chief Medical Officer, Cellecta
Date & Time: Friday, November 3 from 12:00 - 1:30 p.m. and 5:10 - 6:40 p.m in Exhibit Hall B (Poster Hall)
Multi-omic analysis of microsamples of lancet-induced blood drops allows frequent capture and quantitation of numerous metabolites, lipids, cytokines, and proteins. Microsample-based transcriptome profiling would facilitate use of RNA biomarkers n diagnosis and treatment of immunotherapy patients, but a suitable method has not been available. We tested a targeted sequencing protocol for this purpose in human blood microsamples.
Universal synthetic spike-in controls for accurate adaptive immune receptor profiling
Poster number: 154
Presenter: Alex Chenchik, Ph.D., President, Cellecta
Date & Time: Saturday, November 4 from 11:55 a.m. - 1:25 p.m. and 7:00 - 8:30 p.m. in Exhibit Hall B (Poster Hall)
The results of adaptive immune receptor (AIR) repertoire diversity assays can be affected by various biases from differences in conditions in the RT-PCR and NGS sequencing steps. Spike-in synthetic controls can be used as calibration standards to address these biases.
In this study, we synthesized near full-length BCR and TCR constructs that mimic seven different IGH, IGK, IGL, TRB, TRA, TRG and TRD genes. To test our spike-in controls, three sets of variants at different concentrations were added to the RNA samples before the reverse transcription reaction with Cellecta’s DriverMap™ Adaptive Immune Receptor (AIR) Profiling Assay. The DriverMap™ protocol uses reverse gene-specific primers with unique molecular identifiers (UMI), allowing UMI-based correction of amplification biases during data analysis.