Comparative study of serum sample preparation methods in aggregation-based plasmonic sensing.

The use of nanoparticle-based colorimetric methods has received considerable attention in a broad range of clinical and biomedical applications due to their high sensitivity, low cost, extreme simplicity and excellent analytical performance. However, the formation of a protein corona has severely limited the application of nanoparticles (NPs) in clinical samples, which can confer colloidal stability to serum-exposed nanoparticles compared to pristine particles. To address this challenge, dialysis, ultrafiltration and phenol : chloroform : isopentanol extraction methods were compared aiming at facile and routine protein separation methods to eliminate the formation of protein corona on NPs and the development of a sensitive and simple therapeutic drug monitoring (TDM) assay for the detection of aminoglycoside antibiotics in serum.

The study on venom proteins of Lapemis hardwickii by cDNA phage display.

In this study, a cDNA T7 phage display library was constructed from sea snake Lapemis hardwickii venom gland mRNA to analyze the components in venom and find new toxins. All the venom gland cDNA-encoding proteins, including housekeeping proteins and venom proteins were expressed on the surface of bacteriophage T7. This library was then panned with rabbit anti-sea snake venom IgG.

Detection of breast cancer-related antigens through cDNA phage-displayed protein microarray.

High-throughput molecular profiling techniques are helpful in the diagnosis of multifactorial disease. In this study, a cDNA-phage-displayed protein microarray using phage particles spotted directly onto it as sensors was used to detect related antigens in breast tumor sera. cDNA sequences from 17 positive clones were determined, which included some sequences encoding known breast cancer-related antigens and proteins related to other diseases, as well as proteins with unknown functions.

A protein microarray prepared with phage-displayed antibody clones.

Using a large phage antibody library, a protein microarray spotted directly with phage-displayed antibody clones was created to discriminate between recognition profiles of samples from healthy donors and leukemia patients. The protocol for preparing antibody-displaying phage chips was presented. Some conditions such as substrates and blocking buffers were compared and optimized.

Efficient targeted anticoagulant with active RGD motif.

Three anticoagulants combining large peptide recombinant hirudin variants (rHV2-K47) and Arg-Gly-Asp (RGD) motif related to platelet aggregation were generated, i.e. sequences CRFPRGDADPYCE and CNPRGDFRCI were added to the C-terminus of hirudin to obtain RGD-hirudin 1 and 2, respectively, and the sequence RGDSE was inserted between residues 53-54 of hirudin to obtain RGD-hirudin 3.

Construction of a large phage display antibody library by in vitro package and in vivo recombination.

Capacity and diversity are extremely important to the quality of various phage display libraries. In this work, lambda phage-based in vitro package was applied to construct a filamentous phage display antibody library so as to enlarge its capacity and introduce more sequence diversity in the final library. In vivo recombination via Cre recombinase/lox sites was also exploited to create V(H)/V(L) combination diversity based on multivalent package of lambda phage packaging extracts on phagemid DNA concatemers.

Investigation on recombinant hirudin via oral route.

The possibility for oral administration of peptide recombinant hirudin variant (rHV2-K47) as an anticoagulant agent was evaluated in several aspects. The proteolytic properties of rHV2-K47 and its stability during storage were examined by in vitro experiments. Radiolabeled rHV2-K47 was infused into the duodenum of rats and rHV2-K47 absorbed into serum was shown to be intact by electrophoresis pattern.

Molecular cloning, expression and characterization of glutathione S-transferase from Mytilus edulis.

The gene coding for glutathione S-transferase (GST) has been isolated from the Mytilus edulis hepatopancreas. Open reading frame analysis indicated that the M. edulis GST (meGST) gene encodes a protein of 206 amino acid residues with a calculated molecular mass of 23.

Codon optimization of MTS1 and its expression in Escherichia coli.

MTS1, which encodes a protein named p16, is an important gene involved in tumorigenesis. To increase the expression of p16 in Escherichia coli, MTS1 was synthesized de novo by recursive PCR, with codons optimized towards E. coli.

Purification and characterization of a novel glutathione S-transferase from Atactodea striata.

A novel GST isoenzyme was purified from hepatopancreas cytosol of Atactodea striata with a combination of affinity chromatography and reverse-phase HPLC. The molecular weight of the enzyme was determined to be 24 kDa by SDS-PAGE electrophoresis and 48 kDa by gel chromatography, in combination with GST information from literature revealed that the native enzyme was homodimeric with a subunit of M(r) 24 kDa. The purified enzyme, exhibited high activity towards 1-chloro-2,4-dinitrobenzene (CDNB) and 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl).

Prospects of RNA Researches.

No Abstract

Construction and characterization of trifunctional single-chain urokinase-type plasminogen activators.

Two chimeric proteins have been constructed. One consists of four parts: a portion of the low molecular mass single-chain urokinase-type plasminogen activator (scu-PA-32K, residues 144-411), a 15-mer linker sequence, the C-terminal amino-acid sequence (residues 53-65) of hirudin (Hir), and an RGD sequence derived from the leech protein decorsin, i.e.