Experiment system and method for accurate controlling of macromolecular crystallization process

Patent metadata

Field	Value
Publication number	US-11452948-B2
Application number	US-201816349572-A
Country	US
Kind code	B2
Filing date	Dec 18, 2018
Priority date	Dec 27, 2017
Publication date	Sep 27, 2022
Grant date	Sep 27, 2022

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 form 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

The invention claimed is:

1. An experiment-controlling system for accurate controlling of a macromolecular crystallization process, wherein the experiment system is a closed space encircled by a shell and is comprised 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;
wherein 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 platform horizontal moving slot is adjusted through the y-axis horizontal adjusting mechanism, so as to ensure accurate movement of the high-throughput macromolecular crystal cultivation platform;
wherein 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 a volume of a droplet added by the sample injector, and the sample injector height adjusting mechanism is used for controlling the position of the sample injector;
wherein the observing module comprises a high-speed microcamera and a camera adjusting unit, the high-speed microcamera is fixed to the camera adjusting unit, and 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;
wherein the experiment condition control module comprises a temperature and humidity measurement and control apparatus for controlling humidity and temperature required in the closed space;
wherein 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, and the user observing computer is connected with the camera adjusting unit through the data export line; and
wherein 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-controlling system for accurate controlling of the 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; the high-throughput macromolecular crystal cultivation platform is formed by convex structures and concave structures with two different layout forms;
a first layout form of a lattice layout, with the convex structures on one side of the high-throughput macromolecular crystal cultivation platform according to the lattice layout, and with the concave structures on the other side of the high-throughput macromolecular crystal cultivation platform according to the lattice layout; and
a second layout form of a tunnel layout, with the convex structures and the concave structures alternately arranged to form the high-throughput macromolecular crystal cultivation platform with tunnel type.

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

4. An experiment-controlling method for accurate controlling of the macromolecular crystallization process by using the experiment system of claim 1 , comprising the following steps:
(1) preparing a crystal culture solution with a macromolecule crystallization solution and solvent at a temperature of 20-30° C.; disinfecting the high-throughput macromolecular crystal cultivation platform; adding the 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;
(2) adjusting temperature and humidity measurement and control apparatus so that temperature and humidity conditions in a closed space reach values set during crystallization, wherein temperature and humidity throughout an experiment are controlled by the experiment condition control module;
(3) adjusting the x-axis horizontal adjusting mechanism and the y-axis horizontal adjusting mechanism during the experiment, aligning a target site for adding a droplet on the high-throughput macromolecular crystal cultivation platform with the sample injector, adjusting the sample injector height adjusting mechanism according to a height of the high-throughput macromolecular crystal cultivation platform, and rotating a piston propulsion mechanism to extrude the droplet, the volume of which is indicated by graduations on the sample injector;
(4) observing the state of the crystal droplet through the high-speed microcamera of the observing module, converting an observed content into an electrical signal and presenting on a user observing computer through a data export line, and collecting and storing crystallization pictures or videos; and
(5) taking out the high-throughput macromolecular crystal cultivation platform from the platform horizontal moving slot, connecting a pipeline of anti-solvent to the backwash liquor inlet to start washing, washing liquid passing through the backwash liquor flow channel and being collected at the backwash liquor outlet, and backwashing out the crystal to complete operation of high-throughput crystal cultivation.

Assignees

• Univ Dalian Tech

Inventors

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

Classifications

• G01N2021/8477

  Investigating crystals, e.g. liquid crystals · 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

• B01D9/0045

  Washing of crystals, e.g. in wash columns · CPC title

• B01D2009/0086

  Processes or apparatus therefor · CPC title