化学生物学实验技术

Nonviral gene delivery has been gaining considerable attention recently. Although the efficacy of DNA transfection, which is a major concern, is low in nonviral vector-mediated gene transfer compared with viral ones, nonviral vectors are relatively easy to prepare, less immunogenic ...

The construction of safe, efficient, and modifiable synthetic DNA nanoparticles is an emerging technology that has achieved important milestones of success in the past 5 years. Advances in chemical conjugation, purification, and controlled synthesis have allowed researchers to ...

This chapter is a brief description of the state of the art of the field of targeted drug delivery using magnetic implants. It describes the advantages and drawbacks of the use of internal magnets to concentrate magnetic nanoparticles near tumor locations, and the different approaches to this t ...

We developed extremely small functionalized magnetic nanoparticles (MNPs) for use as an in vivo delivery system for pharmaceuticals and biomolecules. We functionalized the MNPs (d = 3 nm) by silanization of amino groups on the particles with (3-aminopropyl)triethoxysilane for sub ...

Examining messenger RNA (mRNA) expression is useful for the determination of cell and tissue conditions. Many methods of determining mRNA expression require total RNA extraction or cell fixation. These processes cause difficulties in examining mRNA expression in single living ce ...

Targeted cancer therapy allows the delivery of therapeutic agents to cancer cells without incurring undesirable side effects on the neighboring healthy tissues. Over the past decade, there has been an increasing interest in the development of advanced cancer therapeutics using tar ...

Reverse transfection from a solid surface has the potential to deliver genes into various types of cell and tissue more effectively than conventional methods of transfection. We present a method for reverse transfection using a gold colloid (GC) as a nanoscaffold by generating nanoclust ...

Single DNA molecule approaches are playing an increasingly central role in the analytical genomic sciences because single molecule techniques intrinsically provide individualized measurements of selected molecules, free from the constraints of bulk techniques, which bl ...

Plastic microfluidic devices are fabricated with an array of pseudo-valves for two-dimensional (2D) protein separation. The devices are made by compression molding; the mold is created by electroplating on a glass master fabricated by photolithography. Each device consists of one ch ...

This chapter introduces the demonstration of specific antibody detection by using a microbead-based assay with quantum dot (QD) fluorescence on a polydimethylsiloxane (PDMS) microfluidic chip. The microfluidic chip is designed to isolate a single microbead where the binding rea ...

Focusing methods are a key component in many miniaturized DNA analysis systems because they enable dilute samples to be concentrated to detectable levels while being simultaneously confined within a specified volume inside the microchannel. In this chapter, we describe a focusing me ...

Solid-state nanopores have been fabricated and used to characterize single DNA and protein molecules. Here we describe the details on how these nanopores were fabricated and characterized, the nanopore sensing system setup, and protocols of using these nanopores to characterize DNA a ...

Force spectroscopy can be applied using nanopores to study charged molecules such as nucleic acids. This technique can be used to study the binding energy of a DNA duplex by threading an anchored single-stranded DNA (ssDNA) probe molecule through a nanopore (having a diameter large enough to ac ...

HPLC-chip/MS is a novel nanoflow analytical technology conducted on a microfabricated chip that allows for highly efficient HPLC separation and superior sensitive MS detection of complex proteomic mixtures. This is possible through on-chip preconcentration and separation wi ...

Confining DNA molecules in a nanofluidic channel, particularly in channels with cross sections comparable to the persistence length of the DNA molecule (about 50 nm), allows the discovery of new biophysical phenomena. This sub-100 nm nanofluidic channel can be used as a novel platform to stu ...

Understanding the safety of newly developed compounds is a key task in each early drug discovery project. In early stages, pharmaceutical companies address this task by using so-called preclinical safety profiling, in which compounds are screened in inexpensive large-scale assays to ...

Chemically similar drugs often bind biologically diverse protein targets, and proteins with similar sequences or structures do not always recognize the same ligands. How can we uncover the pharmacological relationships among proteins, when drugs may bind them in defiance of bioinf ...

Covalent modification of proteins with SUMO (small ubiquitin related modifier) affects many cellular processes like transcription, nuclear transport, DNA repair and cell cycle progression. Although hundreds of SUMO targets have been identified, for several of them the function ...

We previously described the establishment of a binary vector system that allows co-expression of SUMO conjugation enzymes and a target protein of interest, leading to efficient SUMO modification and the production of a large amount of recombinant SUMO-modified proteins in Escheric ...

Ni-NTA affinity chromatography under denaturing conditions has proven to be a powerful method for the isolation of SUMO conjugates from total cell extracts, as it minimizes deconjugation and excludes noncovalent interactions. This chapter describes the use of both His-tagged SUMO a ...