- Although Merrifield invented the solid-phase peptide synthesis science and achieved great success, it used the self-developed synthesis equipment, but it did not bring the peptide synthesis instrument into the market.
- Development of peptide synthesizer
- The design principle of act90 is that the reactor swings left and right around the origin in an upright position, or moves in a circle.
Although Merrifield invented the solid-phase peptide synthesis science and achieved great success, it used the self-developed synthesis equipment, but it did not bring the peptide synthesis instrument into the market.
In 1970, Beckman 990 peptide synthesizer was developed by Beckman company as the first peptide synthesizer for scientific research, which was adopted by laboratories of many universities in the United States.
Almost at the same time, Vega biotechnologies, Inc. developed two economical peptide Synthesizers: Vega’s coupler 1000 and Vega’s coupler 250 (soon Vega’s coupler 296), which combined the subsequent online cutting concept of peptide synthesis into the equipment. All reactors were made of explosion-proof glass to prevent TFA corrosion. At that time, it was called the most economical and applicable peptide synthesis instrument.
Now, Beckman and Vega’s both stopped the R & D and manufacturing of peptide synthesis instrument, and turned to the development industry of chemical synthesis, separation and detection technology and equipment.
Development of peptide synthesizer
The first generation peptide synthesizer is represented by Beckman 990 peptide synthesizer produced by Beckman company and Vega’s 296 peptide synthesizer launched by Vega’s biotechnologies company. It was born in the 1970s.
Although with the improvement and development of production technology, the first generation of peptide synthesizer has been withdrawn from the market. However, many peptide chemistry literatures before 1990 were developed on this experimental equipment. The first generation of peptide synthesizer has great significance for the later research and development of peptide synthesis instrument.
The second generation peptide synthesizer, represented by PS3 peptide synthesizer produced by protein technologies and act peptide synthesizer model 90 by advanced Chemtech, was born in the 1980s. These two devices are also the earliest polypeptide synthesizer still on sale in the market.
The design principle of PS3 is to use nitrogen bubbling reaction mode to stir the reactants, that is, the reactor on the synthesis instrument is fixed, and nitrogen is discharged from the bottom of the reactor through the reactor to the upper part, and the vapor bubbles generated in this process mix the solid and liquid phases. The advantages of this design are simple structure and low cost, but the reaction is relatively mild
1) Sometimes the polypeptide solid carrier will “agglomerate” under the action of static electricity, which makes it unable to fully mix with the liquid phase. In this case, it is necessary to increase the pressure of nitrogen to eliminate the electrostatic effect; however, after the electrostatic effect is eliminated, the pressure should be immediately lowered, otherwise the higher pressure will “blow” the peptide solid carrier above the liquid level of the reactor.
Due to the strong wall adhesion of peptide solid phase carrier, once it is stuck to the reactor surface, it can no longer come down, that is, it can not participate in the reaction. Obviously, the first generation of machines can not automatically make such pressure adjustment, which is an important reason for the “dead angle” of reaction. The dead angle of reaction will reduce the efficiency of peptide synthesis and the purity of peptide, and some even cause the failure of synthesis.
2) After a long time of nitrogen bubbling, the solution will volatilize. After the liquid level is lowered, some polypeptide solid support will stick to the surface of the liquid and can not participate in the reaction again.
3) The nitrogen consumption is large and the operation cost is increased.
The design principle of act90 is that the reactor swings left and right around the origin in an upright position, or moves in a circle.
The peptide synthesizer of act also has the characteristics of mild reaction, that is, the rotation angle and speed can not fully reach the limit of amino acid coupling, and the reaction often takes longer time.
The third generation peptide synthesizer, represented by ABI 433 peptide synthesizer of Applied Biosystems and cs336 of CS bio company, was born in the 1990s.
The design principle of abi433 is that the upper part of the reactor is relatively fixed, while the lower part is rotated rapidly by 360 degrees, which drives the solid-liquid two phases in the reactor to spiral from the bottom to the top of the reactor. In other words, the solution can reach any point in the reactor, and there is no dead corner.
As the stirring rate can reach 1800 rpm, the reaction can be fully completed. Due to the no dead angle stirring method to ensure the purity of peptide synthesis, abi433 peptide synthesizer (the last instrument after its withdrawal from the peptide synthesizer market) still occupies a large proportion in the world. Of course, the price of ABI products is also the highest. Due to the high frequency of components, the solenoid valve will often be damaged. ABI makes 7 solenoid valves into modular design.
If one solenoid valve is broken, the whole module must be replaced, which increases the maintenance cost.
The design principle of cs336 is that the center point of the reactor is the center of the circle, and the stirring speed can reach 180 rpm. At the same time, it adopts the advantages of nitrogen bubbling reaction. Nitrogen blowing is integrated into the reaction method as an optional reaction mode. The high coupling effect of peptide synthesis instrument in scientific research field is fully reflected.
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