Experiment system and method for accurate controlling of macromolecular crystallization process

Patent metadata

Field	Value
Publication number	US-2020108330-A1
Application number	US-201816349572-A
Country	US
Kind code	A1
Filing date	Dec 18, 2018
Priority date	Dec 27, 2017
Publication date	Apr 9, 2020
Grant date	—

Abstract

An experiment system and method for accurate controlling of macromolecular crystallization process. The system has a platform-equipped horizontal moving slot and channel dedicated backwash module, a droplet adding control module, an observing module, a user observation computer system, and an experimental condition control module. A high-precision movement knob of the x-axis platform and the y-axis platform of the system and the accurate position control of a syringe needle are used to ensure that the macromolecular solution can be added into the correct positions of convex or concave. The crystallization induction period of the target crystal faun is determined by the real-time data of the high-speed microcamera, and the crystal cultivation environment is adjusted in real time. This is simple and easy to operate, high in productivity, can be applied to the conventional experimental replication.

First claim

1 . An experiment system for accurate controlling of macromolecular crystallization process, wherein the experiment system is a closed space encircled by a shell at ordinary pressure and is mainly composed of a platform-equipped horizontal moving slot and channel dedicated backwash module, a droplet adding control module, an observing module, a user observation computer system, and an experiment condition control module; the platform-equipped horizontal moving slot and channel dedicated backwash module, the droplet adding control module, the observing module and the experiment condition control module are positioned inside the closed space, and the user observation computer system is positioned outside the closed space;
the platform-equipped horizontal moving slot and channel dedicated backwash module comprises a platform horizontal moving slot, an x-axis horizontal adjusting mechanism, a y-axis horizontal adjusting mechanism and a high-throughput macromolecular crystal cultivation platform; the high-throughput macromolecular crystal cultivation platform is placed on the platform horizontal moving slot; the horizontal position of the platform horizontal moving slot is adjusted through the x-axis horizontal adjusting mechanism and the vertical height of the same is adjusted through the y-axis horizontal adjusting mechanism, so as to ensure accurate movement of the high-throughput macromolecular crystal cultivation platform; the droplet adding control module comprises a sample injector, a piston propulsion adjusting mechanism and a sample injector height adjusting mechanism, the piston propulsion adjusting mechanism is used for accurately controlling the volume of a droplet added by the sample injector, the sample injector height adjusting mechanism is used for controlling the position of the sample injector; the observing module comprises a high-speed microcamera and a camera adjusting unit, the high-speed microcamera is fixed to the camera adjusting unit, a state of crystal inside the droplet on the high-throughput macromolecular crystal cultivation platform is observed by controlling the angle, brightness and magnification factor of the high-speed microcamera; the experiment condition control module comprises a temperature and humidity measurement and control apparatus for controlling humidity and temperature required in the closed space; the user observation computer system comprises a data export line and a user observing computer, and is used as an external extension of the observing system for facilitating the user to observe crystal growth under a camera lens of the high-speed microcamera using computer software; the user observing computer is connected with the camera adjusting unit through the data export line; the high-throughput macromolecular crystal cultivation platform comprises a backwash liquor inlet, a backwash liquor flow channel and a backwash liquor outlet, backwash liquor enters from the backwash liquor inlet, and after washing the crystal in the backwash liquor flow channel, the backwash liquor flows out of the backwash liquor outlet.

2 . The experiment system for accurate controlling of macromolecular crystallization process according to claim 1 , wherein the backwash liquor flow channel is a convex or concave structure of micron scale structure with regular patterns; a crystal platform formed by convex structures and concave structures with different layout forms is selected according to actual needs, and there are two layout forms;
the first layout form is a lattice layout, with the convex structures on one side of the crystal platform according to the lattice layout, and with the concave structures on the other side of the crystal platform according to the lattice layout; the second layout form is a tunnel layout, with the convex structures and the concave structures alternately arranged to form a crystal platform with tunnel type.

3 . The experiment system for accurate controlling of macromolecular crystallization process according to claim 2 , wherein according to the morphology of a target crystal system, the sizes of the convex structures and the concave structures are determined, and the height of the convex structures or the concave structures is 10 μm to 500 μm.

4 . The experiment system for accurate controlling of macromolecular crystallization process according to claim 2 , wherein if the experiment system is used for preparing high-throughput crystal, the crystal platform is designed as the second tunnel layout form to facilitate the washing of anti-solvent and the centralized collection of crystal products, and the connected sample injector is selected to improve the sample introduction efficiency; if the experiment system is used for screening crystal features, the crystal platform is designed as the first lattice layout form to improve the repeatability of crystal screening, and the crystal platform is made of material that does not react with the target crystal system.

5 . The experiment system for accurate controlling of macromolecular crystallization process according to claim 2 , wherein for the temperature and humidity measurement and control apparatus, the control range of humidity is 10% to 100% and the control range of temperature is −50° C. to 200° C.;
the errors of the x-axis horizontal adjusting mechanism, the y-axis horizontal adjusting mechanism and the sample injector height adjusting mechanism are one thousandth per 25.4 mm; and
the magnification factor of the high-speed microcamera is 10 to 600 times.

6 . The experiment system for accurate controlling of macromolecular crystallization process according to claim 4 , wherein for the temperature and humidity measurement and control apparatus, the control range of humidity is 10% to 100% and the control range of temperature is −50° C. to 200° C.;
the errors of the x-axis horizontal adjusting mechanism, the y-axis horizontal adjusting mechanism and the sample injector height adjusting mechanism are one thousandth per 25.4 mm; and
the magnification factor of the high-speed microcamera is 10 to 600 times.

7 . The experiment system for accurate controlling of macromolecular crystallization process according to claim 4 , wherein the connected sample injector improves the sample introduction efficiency through the following operation: adjusting the x-axis horizontal adjusting mechanism and the y-axis horizontal adjusting mechanism, aligning a target site for adding a droplet on the platform with the sample injector of a connected structure, adjusting the sample injector height adjusting mechanism according to the height of the high-throughput macromolecular crystal cultivation platform, when ready, selecting a sample injector control system to extrude the droplet and the volume of which is indicated by the graduations on the sample injector of the connected structure, each sample injector being controlled by the sample injector control system to ensure that the amount of the droplets extruded by each sample injector of the sample injectors of the connected structure are identical, and processing capacities are consistent; the method uses the connected structure to connect a plurality of sample injectors in series or in parallel to increase output efficiency.

8 . An experiment method for accurate controlling of macromolecular crystallization process, comprising the following steps:
(1) placing crystal culture solution prepared with target crystal system and solvent at a temperature of 20-30° C. for standby; disinfecting the crystal platform; adding the prepared crystal culture solution into the sample injector; placing the high-throughput macromolecular crystal cultivation platform on the platform horizontal moving slot, to complete preparation before crystal cultivation

Assignees

• Univ Dalian Tech

Inventors

• Jiang Xiaobin
• Han Mingguang
• He Gaohong
• Li Jin
• Li Xiangcun
• Xiao Wu
• Wu Xuemei

Classifications

• C30B35/00

  Apparatus not otherwise provided for, specially adapted for the growth, production or after-treatment of single crystals or of a homogeneous polycrystalline material with defined structure · CPC title

• B01D9/0077

  Screening for crystallisation conditions or for crystal forms · CPC title

• C30B7/00

  Single-crystal growth from solutions using solvents which are liquid at normal temperature, e.g. aqueous solutions (from molten solvents C30B9/00; by normal or gradient freezing C30B11/00; under a protective fluid C30B27/00) · CPC title

• B01D9/0054

  Use of anti-solvent · CPC title

• C30B29/58 Primary

  Macromolecular compounds · CPC title