Development Of Metalloprotein Separation System Through 3D Printing Technology

As a rapidly developing manufacturing technology,3D printing has appeared in many fields such as construction,machinery and medical treatment with the characteristics of shorter production process,more convenient personalized customization and more effective integrated molding.Along with the development of materials science and the needs of biochemical analysis,it has moved to the laboratory,providing a new choice for the manufacture and improvement of experimental equipment.Based on the multi-function characteristics of the metalloprotein separation system,this paper aims to achieve miniaturization and easy operation.Through the application of assembly in 3D printing,the system is endowed with the properties of multi-materials and multi-precision,which not only makes the original system achieve structural optimization but also realizes functional perfection.Firstly,it summarizes the characteristics of existing methods such as the improvement of printing equipment and the introduction of 4D printing technology,and concludes that the application of assembly in 3D printing is the simplest and fastest way to deal with it.By summarizing the common assembly in traditional manufacturing,such as welding,mortise and tenon joint,thread joint,interference joint and adhesive joint,and combining with the manufacturing characteristics of 3D printing,six kinds of assembly methods were designed respectively,and the parameters were optimized by finite element analysis.Tensile and airtight tests show that within the limit of stress,the sealing effect with leakage rate close to0 can be obtained by screw assembly(ultimate stress is 45 N),interference assembly(ultimate stress is 27 N)and light curing assembly(ultimate stress is 66 N),which provides theoretical basis and design method for the subsequent structural optimization of this study.Then,the separation device was designed by the light curing,thread and interference assembly respectively,and the structure and parameters were optimized by finite element analysis.According to the characteristics of printing method and printing materials,the manufacturing of each part is realized by SLA 3D printing with transparent,white and rubber photosensitive resin as materials.After the assembly operation test,more than 2 hours of tightness test and more than 10 times of gel electrophoresis test,it was found that the gel tube plugged-in metalloprotein separation device was the best choice for this study.At the same time,in order to reduce the difficulty of gelatinization and improve the separation effect of protein gel electrophoresis,the gel tube structure was further adjusted to be"round outside and square inside",the corresponding outer diameter was 7.5 mm and inner diameter was 3.5 mm.After the test,protein bands with a resolution greater than 6σwere obtained.Thus,the closure,operation and separation effect of the metalloprotein separation device were improved.Then,the cooling module,elution module and electrophoresis module of the metalloprotein separation system were designed,printed and tested respectively.In the cooling module,the cooling capacity of the system is above 10 W,and the TECI-12706thermoelectric chip with the size of only 40 mm×40 mm×3.9 mm is selected.Under the control of the single chip microcomputer chip with the volume of less than 1 cm~3,the system operating temperature is kept in the range of-5℃～3℃.Combined with the DS18B20temperature sensor whose monitoring range is-55℃～110℃,the negative feedback adjustment of the cooling module is realized.In the elution module,the flow rate of the system is adjustable within 10μL/min～200μL/min,and the constant flow control with error of 0.3‰is realized by the parallel operation of two pumps with variable volume piston liquid pump as the basic unit.At the same time,open-source design is adopted to realize the mutation-free regulation of flow velocity with the input of program instructions.In the electrophoresis module,the volume of the cooling channel is adjusted to 0.2 L,which is only2/5 of the original design,and a larger heat exchange rate is obtained.The dead volume of the elution interface is optimized to 0.78 mm~3～2.04 mm~3,which is only 1/3 of the original device.The connection between the elution interface and the gel tube was realized by the light curing,thread and interference assembly respectively.Through the finite element analysis and the physical test,the interference structure design meeting the requirements of tightness and easy operation was obtained.In this way,the interaction of cooling module,elution module and electrophoresis module are realized under the operation condition of low temperature and constant current.Finally,according to the shape and size of each component design,the size of the system appearance is 337 mm×308 mm×343 mm,only 2/5 of the original design;At the same time,considering the manufacturing range of the conventional printing equipment,the structure is split and the assembly is realized by the way of secondary light curing.Through the spatial planning of the placement of each module and the on-line gel electrophoresis test of the metalloprotein,the metalloprotein separation system with the characteristics of miniaturization,simple operation,high sensitivity and high resolution was obtained.This study demonstrates the feasibility and universality of assembly in 3D printing.With the help of assembly,the coupling degree between product components is reduced and the advantages of structure,material and precision are integrated.At the same time,combined with 3D printing personalized customization and integrated molding,only 6.72 L photosensitive resin consumption,in 36 hours to get the finished product;Including the cooling and elution device manufacturing,the total cost of the system is controlled within￥30,000 RMB to achieve the design and optimization of the product.