A Novel Reagent for Radioiodine Labeling of New Chemical Entities (NCEs) and Biomolecules

Radioiodine labeling of peptides and proteins is routinely performed by using various oxidizing agents such as Chloramine T, Iodobeads, and Iodogen reagent and radioactive iodide (I−), although some other oxidizing agents were also investigated. The main objective of the present study was to develop and test a novel reagent, inorganic monochloramine (NH2Cl), for radioiodine labeling of new chemical entities and biomolecules which is cost-effective, easy to make and handle, and is selective to label amino acids, peptides, and proteins. The data presented in this report demonstrate that the yields of the non-radioactive iodine labeling reactions using monochloramine are >70% for an amino acid (tyrosine) and a cyclic peptide (cyclo Arg-Gly-Asp-d-Tyr-Lys, cRGDyK). No evidence of the formation of N-chloro derivatives in cRGDyK was observed, suggesting that the reagent is selective in iodinating the tyrosine residue in the biomolecules. The method was successfully translated into radioiodine labeling of amino acid, a peptide, and a protein, Bovine Serum Albumin (BSA).

Efficacy of alcohol/sugar aqueous biphasic system on partition of bovine serum albumin

The efficacy of alcohol/sugar aqueous biphasic (ABS) system on protein extraction was investigated. A model protein, bovine serum albumin (BSA), was adopted to evaluate the effects of types and concentration of phase-forming components, protein concentration, and system pH on the protein partition efficiency. The 1-propanol/maltose ABS exhibited an overall better partition efficiency of BSA to the alcohol-rich top phase. A maximum partition coefficient (K) of 20.01 ± 0.05 and recovery yield (Y) of 95.42% ± 0.01% of BSA were achieved with 35% (w/w) 1-propanol/22% (w/w) maltose ABS at pH 5.0 for 10% (w/w) BSA load. The K and Y of BSA in 1-propanol/maltose ABS was slightly improved with the addition of 3% (w/w) of ionic liquid, 1-butyl-3-methylimidazolium bromide ([Bmim]Br) as the adjuvant that could provide protein stabilizing effect. The Fourier Transform Infrared Spectrum (FTIR) analysis revealed that the protein structure remained unaltered upon the separation process.

Optical multiplexed bioassays on photonic crystals for breast cancer biomarker detection

An optical biosensor for proteomic breast cancer biomarker detection in complex media is presented. Bloch Surface Waves (BSW) excited onto one dimensional photonic crystal (1DPC) were used to probe the interaction of HER2 with three antibody species and an inert protein (Bovine Serum Albumin - BSA). The optical system combines Label-Free readings to track the bioassay real-time development and Fluorescence emission quantification to evaluate the level of specific interaction between the antigen and the antibodies. The results confirm a distinguishable level of affinity between the antibodies and the analyte according to their specificity even at low antibody surface density (about 1173 pg/mm2).

Alteration of water absorption in the THz region traces the onset of fibrillation in proteins

Using terahertz spectroscopy, we established the alteration of the collective hydration of water during the fibrillation process (native → intermediate → fibril) of a model protein bovine serum albumin.

Encapsulation of BSA in hybrid PEG hydrogels: stability and controlled release

Silylated hybrid hydrogels of polyethylene glycol were designed to trap, stabilize and release a model protein (bovine serum albumin). Fine-tuning sol–gel reactions lead to sustained release of BSA over weeks, with good insight of protein stability.

Modulated 3D Cross-Correlation Dynamic Light Scattering Applications for Optical Biosensing and Time-Dependent Monitoring of Nanoparticle-Biofluid Interactions

This paper reviews dynamic light scattering (DLS) of gold nanoparticle-protein interactions for the model protein bovine serum albumin (BSA), as well as in complex biofluids, at the example of mouse serum. DLS data of nanorods of various aspect ratio, of proteins and of mouse serum are discussed in terms of the analysis of their hydrodynamic radii, leading to the distinction of rotational and translational motion as well as to the detection of agglomerates. We address in particular advances obtained by modulated 3D cross correlation dynamic light scattering and recent progress using the CORENN algorithm for analysis of DLS data.

Femtosecond laser programmed artificial musculoskeletal systems

Natural musculoskeletal systems have been widely recognized as an advanced robotic model for designing robust yet flexible microbots. However, the development of artificial musculoskeletal systems at micro-nanoscale currently remains a big challenge, since it requires precise assembly of two or more materials of distinct properties into complex 3D micro/nanostructures. In this study, we report femtosecond laser programmed artificial musculoskeletal systems for prototyping 3D microbots, using relatively stiff SU-8 as the skeleton and pH-responsive protein (bovine serum albumin, BSA) as the smart muscle. To realize the programmable integration of the two materials into a 3D configuration, a successive on-chip two-photon polymerization (TPP) strategy that enables structuring two photosensitive materials sequentially within a predesigned configuration was proposed. As a proof-of-concept, we demonstrate a pH-responsive spider microbot and a 3D smart micro-gripper that enables controllable grabbing and releasing. Our strategy provides a universal protocol for directly printing 3D microbots composed of multiple materials.

Agarose-Chitosan Based Hydrogel Waveguide Matrix: Comparison Synthesis and Performance for Optical Leaky Waveguide (OLW) Biosensor

In this work, a hydrogel-based optical leaky waveguide (OLW) biosensor was developed to detect biomolecules using changes in refractive index (Dn). Hydrogels formed from chitosan and agarose were synthesised to be used as a waveguiding thin film in OLW biosensor. The concentration and thickness of the hydrogels defined by the spin coater speeds and time were optimised to produce a device chip with a single waveguide moded operation of OLW biosensor. The cladding layer was fabricated using 9.5nm titanium coated glass substrate for metal-clad (MCLW) and addition of dye particles (reactive blue 4) for dye-clad (DCLW) format. For the performance of the fabricated device for detection of Dn of a glycerol solution, the highest sensitivity of detection was obtained from MCLW chip made of 2% w/v of chitosan-agarose hydrogel spun at 6000 rpm with LOD: 4.28 x 10–6 RIU, while for DCLW format, the LOD measured at 7.46 x 10–6 RIU. The performance of the device to monitor the protein (bovine serum albumin; BSA and anti-BSA) interaction, the binding affinity in respect of change in peak angle (DӨ°) of chitosan matrix to the protein was measured at DӨ°~(6.2 ± 0.5°)x10–6.

The Effect of Fatty Acids and BSA Purity on Synthesis and Properties of Fluorescent Gold Nanoclusters

Fluorescent gold nanoclusters (AuNCs) are envisaged as a novel type of fluorophores. This work reports on the first comparative study investigating the effect of presence/absence/abundance of fatty acids (namely palmitic acid, PA) or other substances (like glycoproteins and globulins) in the protein (bovine serum albumin, BSA) on synthesis and properties of the final AuNCs. The most popular template (BSA) and microwave (MW)-assisted synthesis of AuNCs have been intentionally chosen. Our results clearly demonstrate that the fluorescent characteristics (i.e., fluorescence lifetime and quantum yield) are affected by the fatty acids and/or other substances. Importantly, the as-prepared AuNCs are biocompatible, as determined by Alamar Blue assay performed on Hep G2 cell line.