Antibodies are critical reagents to detect and characterize human proteins. The biomedical research community aspires to have at least one potent and selective renewable antibody for each human protein, and for each of the common applications. To quantify the potential to achieve this goal with commercial reagent antibodies, we applied a standardized characterization approach to assess the performance of 614 commercial antibodies for 65 neuroscience-related proteins in Western blot, immunoprecipitation, and immunofluorescence applications, using isogenic knockout cells as controls. By carrying out side-by-side comparisons of the antibodies, we demonstrate that: i) for each application tested, 61% to 89% of this protein set was covered by at least one high-performing antibody with 54% to 77% covered by at least one high-performing renewable antibody, suggesting that coverage of human proteins by commercial antibodies is significant; ii) on average recombinant antibodies performed better than monoclonal or polyclonal antibodies; iii) hundreds of underperforming antibodies have been used in published articles. Encouragingly, of the commercial antibodies that did not perform as expected, 18% were removed from the market by manufacturers and 37% had alterations made to their recommended usage. This work suggests that the first step toward achieving coverage of the human proteome with selective renewable antibodies is to mine the existing commercial antibody repertoire, and then use this knowledge to focus new renewable antibody generation efforts.
Macromolecular
bottlebrushes (MBs) are emerging as an attractive
polymer architecture for biomedical applications. Herein, synthesis
and characterization of sulfoxide-containing water-soluble MBs with
poly(2-(methylsulfinyl)ethyl acrylate) (PMSEA) side chains with varying
grafting densities are reported. Highly hydrophilic PMSEA side chains
were prepared by grafting-from poly(2-bromoisobutyryloxyethyl methacrylate)
(PBiBEM) using photoATRP with either CuBr2/TPMA (tris(2-pyridylmethyl)amine),
CuBr2/TPMA*3 (tris([(4-methoxy-2,5-dimethyl)-2-pyridyl]methyl)amine),
or CuBr2/Me6TREN (tris(2-dimethylaminoethyl)amine),
which were photochemically reduced to the appropriate amount of activators
CuBr/ligand to start atom transfer radical polymerization (ATRP).
Kinetic experiments showed that the polymerization was the fastest
using CuBr2/Me6TREN at 6-fold excess of ligand
to Cu. Additionally, the increase of ligand concentration resulted
in a faster reaction with CuBr2/Me6TREN. A series
of PMSEA MBs with grafting densities of 30%, 50%, and 100% was synthesized.
The PMSEA MBs exhibited tunable hydrodynamic lubrication properties
and low cytotoxicity against different types of cells. PMSEA MBs reveal
rheological properties characteristic for nonentangled polymer melts.
Densely grafted PMSEA MBs showed a nonlinear relationship between
the coefficient of friction and the applied force.
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