ONCYTE porous nitrocellulose (PNC) film slides are a 3-D substrate very well suited for protein microarrays due to its high binding capacity and ability to retain proteins’ conformation and antigenic sites. PNC slides have proven extremely valuable for Reverse-Phase Protein Arrays (RPPAs) in the field of cancer research.In recent years, interest in further improving the quantitation of RPPAs has resulted in the use of quantum nanocrystals as detector molecules on ONCYTE AVID PNC. Here, Wang, et al. from UT Southwestern Medical Center applied quantum nanocrystals and confocal laser scanning to RPPAs for studying the dynamics of the oncogenic mutant EGFRvIII signaling network in glioblastoma with a 61 antibody biomarker panel (1). This non-amplified detection system resulted in quantitation down to 0.1 pg protein, comparable to standard amplified RPPA methods and 1000-fold more sensitive than with Western blot methods. Vast improvements were noted in the linear dynamic range which spanned nearly 3 orders of magnitude compared to non-linear results obtained with enzymatic amplification and resulting in more accurate quantitation. In their studies, the investigators found the expected activation of pERK, Akt, Src, and JNK after tetracycline treatment. But interestingly, pSrc and pJNK were also activated at much higher levels. The results suggest that multi-target treatment of glioblastoma targeting EGFRvIII and Src/JNK kinases in future studies may be very efficacious. The full publication can be found here:
- PMID: 21917185 [PubMed]
PMCID: PMC3182966