Single cell analysis of heterogeneous circulating tumor cells (CTCs), by which the genomic profiles of rare single CTCs are connected to the clinical status of cancer patients, is crucial for understanding cancer metastasis and the clinical impact on patients. However, the heterogeneity in genotypes and phenotypes and rarity of CTCs have limited extensive single CTC genome research, further hindering clinical investigation. Despite recent efforts to build platforms that separate CTCs, the investigation on CTCs is difficult due to the lack of a retrieval process at the single cell level. In this study, laser‐induced isolation of microstructures on an optomechanically‐transferrable‐chip and sequencing (LIMO‐seq) is applied for whole genome sequencing of single CTCs. Also, the whole genome sequences and the molecular profiles of the isolated single cells from the whole blood of a breast cancer patient are analyzed.
In antibody discovery, in-depth analysis of an antibody library and high-throughput retrieval of clones in the library are crucial to identifying and exploiting rare clones with different properties. However, existing methods have technical limitations, such as low process throughput from the laborious cloning process and waste of the phenotypic screening capacity from unnecessary repetitive tests on the dominant clones. To overcome the limitations, we developed a new high-throughput platform for the identification and retrieval of clones in the library, TrueRepertoire™. This new platform provides highly accurate sequences of the clones with linkage information between heavy and light chains of the antibody fragment. Additionally, the physical DNA of clones can be retrieved in high throughput based on the sequence information. We validated the high accuracy of the sequences and demonstrated that there is no platform-specific bias. Moreover, the applicability of TrueRepertoire™ was demonstrated by a phage-displayed single-chain variable fragment library targeting human hepatocyte growth factor protein.
Microparticle‐based drug delivery is a promising technology for small volume bioassay platforms. The general utilization of the drug‐loaded microparticles in the in vitro bioassay platforms requires the drug loading method, which should impregnate the general drug types (e.g., water insoluble) with high payload into the variously designed microparticles. Loading the drug into the prefabricated microparticles using solvent evaporation satisfies the requirement. However, similar to the “coffee‐ring effect,” drugs are loaded in a seriously nonuniform manner, caused by the capillary flow during the evaporation process. Here, it is presented that the freeze‐drying is an efficient way to load uniform and high amount of the drug into the prefabricated microparticles. It is demonstrated that freezing solvent can block the capillary flow during the solvent removal process, improving the loading uniformity. The delivered amount of drugs is linearly proportional to the initial loading amount of drugs. Also, this drug loading method is shown to be applied to the various drug types and the prefabricated microparticles with different properties. Considering many challenges to suppress the “coffee‐ring effect” that induces nonuniform impregnation/deposition, the proposed concept can be applied not only for microparticle‐based drug delivery but also for uniform coating applications (e.g., thin‐film coating, DNA/protein microarray).
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