In 1965, Chinese scientists synthesized bovine insulin for the first time in the world, opening a new era of life chemistry research. Thanks to the continuous efforts of scientific researchers in the past decades, great progress has been made in the artificial chemical synthesis of proteins. Compared to natural biosynthesis, chemical synthesis can create man-made proteins with a variety of precisely controlled and non-natural structures, bringing new opportunities for developing protein tools and protein products that meet our needs.

Recently, researchers have made new achievements in the field of chemical synthesis of proteins, and applied related chromatographic column products from Welch Materials. Come and enjoy the feast of scientific research!

Chemical synthesis of large mirror polymerase and realization of mirror DNA information storage

It is reported that DNA in its natural state has undergone delicate evolution to store genetic information. The chiral inverted-stranded L-DNA has the same information ability, is resistant to biodegradation, and can be used as a robust bioorthogonal information library. In a new study, researchers from Tsing Zhu’s group at the School of Life at Tsinghua University chemically synthesized a 90kda high-fidelity mirror Pfu DNA polymerase, which can precisely assemble a kilobase-sized mirror gene. The large-scale mirror protein total chemical synthesis strategy and the kilobase-length mirror gene assembly technology used in this experiment for the first time solve the problem of preparing large mirror biomolecules that have long restricted the development of mirror biology. The research achievement is titled “Bioorthogonal information storage in L-DNA with a high-fidelity mirror-image Pfu DNA polymerase” (Bioorthogonal information storage in L-DNA with a high-fidelity mirror-image Pfu DNA polymerase). Published in the journal Nature Biotechnology on May 29.

Research results at a glance

The researchers used polymerases to encode in L-DNA an 1860 quote by Louis Pasteur that first suggested a mirror world in biology. To break through the protein size limitation of full chemical synthesis, the research team achieved chiral steganography by embedding chimeric D-DNA/L-DNA key molecules into the D-DNA repository. The team divided the Pfu DNA polymerase with a full length of 775 amino acids into two fragments with a length of 467 amino acids and 308 amino acids, which were synthesized separately. After mixing them, they were renatured together to make them correctly folded into a fully functional 90 kDa. The high-fidelity mirror Pfu DNA polymerase is the largest fully chemically synthesized protein reported so far; the researchers also used the high-fidelity mirror polymerase to assemble a mirror 16S ribosomal RNA gene up to 1.5 kb, which is the longest reported so far. Mirror DNA. In addition, they found that trace amounts of L-DNA barcodes preserved in natural conditions (local pond water) could still be amplified and sequenced within 1 year, while D-DNA barcodes under the same conditions could no longer be amplified after 1 day. increase. There is only one reason behind it: their chirality is different.

In the study, the research group used Ultisil® XB-C4 (4.6250mm, 5μm) to monitor the progress of the reaction and check the purity of the peptide product. At the same time, use preparative columns Ultisil® XB-C4 and C18 (21.2250mm, 5μm or 10*250mm, 5μm) to separate and prepare crude peptide fragments and ligation products.

Total chemical synthesis of disulfide-rich proteins

Disulfide-rich proteins are useful drug or tool molecules in biomedical research, but their synthesis is complicated by folding difficulties. In view of this, researchers such as Prof. Liu Lei from Tsinghua University and Prof. Zheng Jishen from University of Science and Technology of China used a removable O-linked β-N-acetylglucosamine strategy to achieve a correctly folded disulfide-rich protein. The research results are titled “Total Chemical Synthesis of Correctly Folded Disulfide-Rich Proteins Using a Removable O-Linked β-N-Acetylglucosamine Strategy” and published in the journal JACS on January 3, 2022.

Research results at a glance

The researchers describe a removable glycosylation modification (RGM) strategy that accelerates the chemical synthesis of properly folded proteins with multiple or even interchain disulfide bonds. During the experiment, Ultisil® XB-C4 (120Å or 300Å, 250mm×4.6mm, 5μm) was used to monitor the protein synthesis reaction, and it was successfully prepared with semi-preparative columns Ultisil® XB-C4 and C18 (300Å, 250mm×10mm, 5μm) get the target protein. This strategy involves the introduction of a simple O-linked β-N-acetylglucosamine (O-GlcNAc) group at the Ser/Thr site, which effectively facilitates the folding of disulfide-rich proteins by stabilizing their folding intermediates . After folding, the O-GlcNAc group can be efficiently removed with β-N-acetylglucosaminidase (OGA), resulting in a correctly folded protein. Using this strategy, the research group accomplished the synthesis of properly folded hepcidin, an iron-regulating hormone that contains four sets of disulfide bonds. For the first time, the researchers achieved total synthesis of properly folded interleukin 5 (IL-5), a 26-kDa homodimeric cytokine responsible for eosinophil growth and differentiation.

“If you want to do a good job, you must first sharpen your tools.” Welch Materials specializes in the development of biological samples such as peptides and proteins, and has launched the Welch biological sample analysis method development kit to help cutting-edge scientific research and daily production analysis and preparation.

P/NProduct Description
00201-33043Ultisil® XB-C18,5μm,300Å,4.6×250mm
00208-33043Ultisil® LP-C18,5μm,300Å,4.6×250mm
00216-33043Ultisil® XB-C4,5μm,300Å,4.6×250mm
00202-33043Ultisil® XB-C8,5μm,300Å,4.6×250mm
00209-33043Ultisil® LP-C8, 5μm, 300Å, 4.6×250mm
00608-31012Blossmate® C4,3.5μm,450Å,2.1×100mm
00608-31010Blossmate® C4,3.5μm,450Å,2.1×50mm

● Method development for proteins, peptides or other macromolecules. In order to better interact with the bonded phase, a filler with a large pore size (300Å or 450Å) is required.

● Different selective bonding phases with different retention capabilities, meet the retention and separation of proteins and peptides of various molecular sizes.

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