CN1368639A - Protein chip - Google Patents

Protein chip Download PDF

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Publication number
CN1368639A
CN1368639A CN 01103524 CN01103524A CN1368639A CN 1368639 A CN1368639 A CN 1368639A CN 01103524 CN01103524 CN 01103524 CN 01103524 A CN01103524 A CN 01103524A CN 1368639 A CN1368639 A CN 1368639A
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CN
China
Prior art keywords
host material
protein
protein chip
membrane
micron
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Granted
Application number
CN 01103524
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Chinese (zh)
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CN1249438C (en
Inventor
徐荣臻
郑树
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Individual
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Individual
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Priority to CN 01103524 priority Critical patent/CN1249438C/en
Publication of CN1368639A publication Critical patent/CN1368639A/en
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Publication of CN1249438C publication Critical patent/CN1249438C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

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  • Separation Using Semi-Permeable Membranes (AREA)
  • Manufacture Of Porous Articles, And Recovery And Treatment Of Waste Products (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

A protein chip includes substrate material and capture-probe. The said substrate material is a porous film of cellulose. Optimal range of aperture size of the porous film is form 0.2-0.6 micro, and the optimal range is 0.2-0.45 micro. The invention possesses high ability of adsorbing protein that the upper limit of adsorbing protein can reach 80 microgrammes of bovine serum albumin per square centimeter. Moreover, adsorbed protein is distributed on surface of the film evenly with high sensibility and good stability. The invention makes the reaction stand in high speed and dynamic state so as to reduce the reaction time greatly. The equilibrium time of the reaction is only below 5 minutes.

Description

A kind of protein chip
The present invention relates to detect technical field with analyzing gene expression product protein and polypeptide.
Theoretically, advantages such as protein chip technology has rapidly and efficiently, sensitive special, easy and simple to handle, cheap practicality, prospect is widely used in fields such as numerous life subjects such as preventive medicine, preclinical medicine, clinical medicine.In protein chip technology, chip matrix is one of core component, and seeking and develop the protein chip host material with superperformance is one of problem anxious to be solved in current this field.At present, the protein chip of developing adopts the host material of glass sheet as chip more, and there are two serious defectives in this host material: (1) protein adsorption capacity is poor, directly influences the sensitivity and the stability of protein chip technology; (2) biomolecular reaction must carry out under solid state shape, and reaction velocity is very slow, needs 2 hours just can reach molecular balance.
The protein chip that the purpose of this invention is to provide a kind of high protein adsorption capacity, sensitivity height and good stability.
Further aim of the present invention is to state in realization on the basis of purpose, provides a kind of and can make the fireballing protein chip of biomolecular reaction.
For realizing purpose of the present invention, the present invention by the following technical solutions: it comprises host material and is located at capture probe on the host material that described host material is meant the porous cellulose film of aperture below 0.6 micron.
For realizing further aim of the present invention, when adopting above technical scheme, the present invention adopts following further technical scheme again: the preferable range in described cellulose membrane aperture is the 0.2-0.6 micron.
The optimized scope in above-mentioned cellulose membrane aperture is the 0.2-0.45 micron.
The preferably polyvinylidene fluoride blotting membrane (being pvdf membrane) of above-mentioned cellulose membrane.。
The preferably nitrocellulose filter of above-mentioned cellulose membrane.
Described capture probe be with the micro-array spray printing on the porous cellulose film.
Owing to adopt technique scheme, cellulose membrane is compared with glass sheet, not only has very high protein adsorption capacity, can reach every square centimeter 80 microgram bovine serum albumin(BSA) to the absorption of proteins upper limit, and the protein that is adsorbed is very even in the film surface distributed, with this protein chip sensitivity height and good stability as host material.What have further effect is, cellulose membrane also can be made vesicular, the 0.2-0.6 micron is adopted in the aperture, especially 0.2-0.45 micron, can make the easy on and off motion of the biomolecule film in reaction chamber that participates in reaction, reaction is in high speed dynamic response state, has shortened the reaction time greatly, reacting balance time only needs also to be of value to the sensitivity and the stability of protein chip simultaneously below 5 minutes.
Below in conjunction with drawings and Examples the present invention is further described.
Fig. 1 is an enlarged diagram of the present invention.
Embodiment 1
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.3 micron in aperture, and this cellulose membrane is a pvdf membrane.
Embodiment 2
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.35 micron in aperture, and this cellulose membrane is a pvdf membrane.
Embodiment 3
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.25 micron in aperture, and this cellulose membrane is a nitrocellulose filter.
Embodiment 4
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.4 micron in aperture, and this cellulose membrane is a nitrocellulose filter.
Embodiment 5
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.2 micron in aperture, and this cellulose membrane is a cellulose acetate membrane.
Embodiment 6
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.45 micron in aperture, and this cellulose membrane is a nitric acid acetic acid cellulose mixture film.
Embodiment 7
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.15 micron in aperture, and this cellulose membrane is a nitrocellulose filter.
Embodiment 8
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.5 micron in aperture, and this cellulose membrane is a pvdf membrane.
Embodiment 9
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.1 micron in aperture, and this cellulose membrane is a nitrocellulose filter.
Embodiment 10
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.55 micron in aperture, and this cellulose membrane is a pvdf membrane.
Embodiment 11
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.05 micron in aperture, and this cellulose membrane is a cellulose acetate membrane.
Embodiment 12
The present invention includes host material 1 and be located at capture probe 2 on the host material, described host material is the porous cellulose film in 0.6 micron in aperture, and this cellulose membrane is a nitric acid acetic acid cellulose mixture film.
The present invention can adopt following method manufacturing: (1) cuts the porous cellulose film in required size and required aperture with stainless steel aseptic operation blade, as 1 square centimeter 0.2 micron pore size pvdf membrane, immerses in the absolute methanol 5 minutes; (2) this film was changed in the damping fluid of 0.1M PBS, PH7.4 over to submergence 5 minutes with aseptic blunt-ended forceps, take out and put the room temperature airing; (3) with micro-spray printing equipment with capture probe with microarray form spray printing to pvdf membrane, every 0.5 microlitre, room temperature airing; (4) with the 0.1MPBS that contains 1% bovine serum albumin(BSA), the damping fluid of PH7.4,37 ℃ of sealings 1 hour; (5) 80 ℃ of vacuum drying were the protein chip for preparing in 2 hours.

Claims (6)

1, a kind of protein chip comprises host material and is located at capture probe on the host material, it is characterized in that described host material is meant the porous cellulose film of aperture below 0.6 micron.
2, a kind of protein chip as claimed in claim 1, the preferable range that it is characterized in that described cellulose membrane aperture is the 0.2-0.6 micron.
3, a kind of protein chip as claimed in claim 1, the optimized scope that it is characterized in that described cellulose membrane aperture is the 0.2-0.45 micron.
4, a kind of protein chip as claimed in claim 1 is characterized in that the preferably polyvinylidene fluoride blotting membrane (being pvdf membrane) of described cellulose membrane.
5, a kind of protein chip as claimed in claim 1 is characterized in that the preferably nitrocellulose filter of described cellulose membrane.
6, as claim 1,2,3,4 or 5 described a kind of protein chips, it is characterized in that described capture probe be with the micro-array spray printing on the porous cellulose film.
CN 01103524 2001-02-02 2001-02-02 Protein chip Expired - Fee Related CN1249438C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN 01103524 CN1249438C (en) 2001-02-02 2001-02-02 Protein chip

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN 01103524 CN1249438C (en) 2001-02-02 2001-02-02 Protein chip

Publications (2)

Publication Number Publication Date
CN1368639A true CN1368639A (en) 2002-09-11
CN1249438C CN1249438C (en) 2006-04-05

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Family Applications (1)

Application Number Title Priority Date Filing Date
CN 01103524 Expired - Fee Related CN1249438C (en) 2001-02-02 2001-02-02 Protein chip

Country Status (1)

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CN (1) CN1249438C (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100386627C (en) * 2001-07-03 2008-05-07 包刚 Filtration-based microarray chip
CN101858902A (en) * 2010-06-01 2010-10-13 中国科学院广州地球化学研究所 Soil alteration carbonate measuring method integrating carbon isotope ratio
CN102667479A (en) * 2009-11-24 2012-09-12 韩国生命工学研究院 Membrane biosensor to which a porous film is attached, and a method for measuring immune reactions or enzyme reactions employing the same
CN101477117B (en) * 2009-01-16 2012-12-19 中国农业科学院哈尔滨兽医研究所 Visible protein chip for detecting poultry disease serum antibody, its preparation method and application

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100386627C (en) * 2001-07-03 2008-05-07 包刚 Filtration-based microarray chip
CN101477117B (en) * 2009-01-16 2012-12-19 中国农业科学院哈尔滨兽医研究所 Visible protein chip for detecting poultry disease serum antibody, its preparation method and application
CN102667479A (en) * 2009-11-24 2012-09-12 韩国生命工学研究院 Membrane biosensor to which a porous film is attached, and a method for measuring immune reactions or enzyme reactions employing the same
CN101858902A (en) * 2010-06-01 2010-10-13 中国科学院广州地球化学研究所 Soil alteration carbonate measuring method integrating carbon isotope ratio

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CN1249438C (en) 2006-04-05

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