CN1220059C

CN1220059C - Method for producing biological protein molecular chips

Info

Publication number: CN1220059C
Application number: CN 200310107947
Authority: CN
Inventors: 焦正; 吴明红; 施利毅; 雷红
Original assignee: University of Shanghai for Science and Technology
Current assignee: Shanghai University; University of Shanghai for Science and Technology
Priority date: 2003-10-16
Filing date: 2003-10-16
Publication date: 2005-09-21
Anticipated expiration: 2023-10-16
Also published as: CN1529169A

Abstract

The present invention relates to a method for producing biological protein chips, substantially an active assembly method for protein molecules on silicon surfaces. The method can be applied to the production of a DNA chip and a protein chip, and belongs to the technical field of biological sensing. The method of the present invention is characterized in that firstly, a silicon chip is washed and thermally oxidized by a standard semiconductor technology; then, octadecy l trichlorosilane is used for processing a silicon surface; a nano figure is etched on the surface of the silicon chip by a nano etching technique of an atomic force microscope (AFM); subsequently, gamma-amino-propyl-triethylsilane is used for processing the silicon surface; finally, the silicon chip in the position of the processed nano figure is immersed in protein solution so that protein molecules are fixed on the surface of the silicon chip in the position of a non-etching figure; the etched surface of the silicon chip does not have the protein molecules. The method of the present invention has the advantages of effectively ensuring the activity of the protein molecules and enhancing combined firm degree, and also has a super-high integration level of the protein molecules.

Description

The manufacture method of bioprotein molecule chip

Technical field

The present invention relates to a kind of manufacture method of bioprotein molecule chip, come down to the active assembly method of a kind of protein molecule at silicon face, this method can be applied to the manufacturing of DNA chip and protein-chip, belongs to the biosensor technique field.

Background technology

DNA chip or protein-chip are on the surface area of a very little geometric scale, integrated multiple DNA or activity of proteins.Only use the sampling of the biology (physiology) of trace can detect and study different molecules simultaneously, comprise interaction and expression of gene between the molecule, the condition that obtains group of molecules under the various conditions changes, thereby can obtain the rule of vital movement, biomolecule chip and biomolecule identification and detection technique, the present cutting edge technology that has become the 21 century biomedical engineering.

As everyone knows, environment is very responsive to external world owing to biomolecule, is easy to lose activity.The key link of protein-chip manufacturing is that protein molecule is transferred on the silicon substrate, has not only required high integration but also must guarantee the activity of protein molecule.Traditional photoetching technique, at the figure that the silicon face etching is processed, its minimum dimension only can improve hundreds of nanometers to several microns, and its integrated level is also not very high, and still needing further, 1 to 2 order of magnitude of raising could satisfy current technical requirement.

Summary of the invention

The object of the present invention is to provide the active assembly method of a kind of protein molecule, can prepare the protein-chip of superelevation integrated level at silicon face.Another object of the present invention is by specific lithographic technique, makes the protein-chip with superelevation integrated level.

The manufacture method of a kind of bioprotein molecule chip of the present invention is characterized in that having following each processing step:

A. at first adopt the semiconductor technology of standard to clean and the thermal oxide silicon chip;

B. above-mentioned pre-service is good silicon chip immerses in the octadecyl trichlorosilane, soaks certain hour and makes silicon chip surface have octadecyl, and octadecyl has repellency to protein molecule;

C. adopt the silicon chip surface etching nano graph of atomic force microscope nanometer lithographic technique in above-mentioned processing;

The method of etching is: apply voltage between atomic force microscope probe and silicon face, traveling probe forms the figure of tens nano-widths at silicon face, and the octadecyl that makes the figure position that is etched is destroyed and remove;

D. the above-mentioned silicon chip that processes is immersed in gamma-amino-propyl group-triethyl silicane, the ruined nano graph of octadecyl surface has amino on the silicon chip, and this amino has compatibility to protein molecule;

E. the above-mentioned silicon chip that has nano graph is immersed in the protein molecule solution, protein molecule is fixed on the silicon chip surface of etching graph position.

Protein-chip manufacture method of the present invention, the method for using than present tradition has following some advantage and effect:

1. method of the present invention, the figure of its etching than low 1 to 2 order of magnitude of present technology, thereby make the integrated level of protein-chip improve 1 to 2 order of magnitude generally tens nanometers.

2. method of the present invention can effectively guarantee the activity of protein molecule, and can improve the firm degree of protein molecule combination.

3. traditional photoetching technique is difficult to reach nano level detecting unit, and integrated level is difficult to break through, and the present invention can reach nano level detecting unit, and integrated level can reach the superelevation degree.

Description of drawings

Fig. 1 is the process flow diagram that the reactive protein molecule is fixed at silicon face in the inventive method.

Embodiment

After now in conjunction with the accompanying drawings specific embodiments of the invention being described in.

Embodiment 1: concrete processing step of the present invention is as follows:

A. at first, adopt standard semiconductor technology to clean and the thermal oxide silicon chip; Silicon chip surface after cleaning and the thermal oxide has the silicon dioxide layer of one deck 500 nanometers;

B. above-mentioned pre-service is good silicon chip immersed in the octadecyl trichlorosilane (OTS) 10 minutes, made silicon chip surface have octadecyl, and octadecyl has repellency to protein molecule;

C. adopt the silicon chip surface etching nano graph of atomic force microscope (AFM) nanometer etching technology in above-mentioned processing; The method of etching is: apply voltage between atomic force microscope probe and silicon face, make the atom under the probe be oxidized to silicon dioxide; Traveling probe is etched into the quadrate array of the length of side 50 nanometers, and surface, the figure position that is etched octadecyl is removed.

D. (among the γ-APTES), soaked 10 minutes, make that the ruined nano graph of octadecyl surface has amino on the silicon chip, this amino has compatibility to protein molecule the above-mentioned silicon chip that processes to be immersed gamma-amino-propyl group-triethyl silicane;

E. the above-mentioned silicon chip that has nano graph is immersed 20% ferritin molecular solution, protein molecule is fixed on have the silicon chip surface of amino nano graph position, form the ferritin nano-dot matrix of the length of side 50 nanometers at silicon chip surface.

Embodiment two: concrete processing step of the present invention is as follows:

A. at first, adopt standard semiconductor technology to clean and the thermal oxide silicon chip; Silicon chip surface after cleaning and the thermal oxide has the silicon dioxide layer of one deck 800 nanometers;

C. adopt the silicon chip surface etching nano graph of atomic force microscope (AFM) nanometer etching technology in above-mentioned processing; The method of etching is: apply voltage between atomic force microscope probe and silicon face, make the atom under the probe be oxidized to silicon dioxide; Traveling probe is etched into radius and is respectively 200 nanometers, 400 nanometers, and live width is the concentric circles of 50 nanometers, and the octadecyl on the surface, figure position that is etched is removed by broken ring.

D. (among the γ-APTES), soaked 20 minutes, make that the ruined nano graph of octadecyl surface has amino on the silicon chip, this amino has compatibility to protein molecule the above-mentioned silicon chip that processes to be immersed gamma-amino-propyl group-triethyl silicane;

E. the above-mentioned silicon chip that has nano graph is immersed 20% ferritin molecular solution, protein molecule is fixed on have the silicon chip surface of amino nano graph position, form radius at silicon chip surface and be respectively 200 nanometers, 400 nanometers, live width is the concentric circles ferritin molecular modeling of 50 nanometers.

Claims

1. the manufacture method of a bioprotein molecule chip is characterized in that having following each processing step:

C. adopt the silicon chip surface etching nano graph of atomic force microscope nanometer lithographic technique in above-mentioned processing; The method of etching is: apply voltage between atomic force microscope probe and silicon face, traveling probe forms the figure of tens nano-widths at silicon face, and the octadecyl that makes the figure position that is etched is destroyed and remove;