+

WO2003099999A3 - Generation and selection of protein library in silico - Google Patents

Generation and selection of protein library in silico Download PDF

Info

Publication number
WO2003099999A3
WO2003099999A3 PCT/US2003/016037 US0316037W WO03099999A3 WO 2003099999 A3 WO2003099999 A3 WO 2003099999A3 US 0316037 W US0316037 W US 0316037W WO 03099999 A3 WO03099999 A3 WO 03099999A3
Authority
WO
WIPO (PCT)
Prior art keywords
library
lead
sequence
protein
proteins
Prior art date
Application number
PCT/US2003/016037
Other languages
French (fr)
Other versions
WO2003099999A2 (en
Inventor
Peizhi Luo
Mark Hsieh
Pingyu Zhong
Caili Wang
Yicheng Cao
Shengjiang Liu
Original Assignee
Abmaxis Inc
Peizhi Luo
Mark Hsieh
Pingyu Zhong
Caili Wang
Yicheng Cao
Shengjiang Liu
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US10/153,176 external-priority patent/US20030022240A1/en
Priority claimed from US10/153,159 external-priority patent/US7117096B2/en
Application filed by Abmaxis Inc, Peizhi Luo, Mark Hsieh, Pingyu Zhong, Caili Wang, Yicheng Cao, Shengjiang Liu filed Critical Abmaxis Inc
Priority to CN038173603A priority Critical patent/CN1672160B/en
Priority to AU2003248548A priority patent/AU2003248548B2/en
Priority to JP2004508241A priority patent/JP2005526518A/en
Priority to CA002485732A priority patent/CA2485732A1/en
Priority to EP03755415A priority patent/EP1514216A4/en
Publication of WO2003099999A2 publication Critical patent/WO2003099999A2/en
Publication of WO2003099999A3 publication Critical patent/WO2003099999A3/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
    • G16B20/00—ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations
    • G16B20/30—Detection of binding sites or motifs
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
    • G16B30/00—ICT specially adapted for sequence analysis involving nucleotides or amino acids
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
    • G16B30/00—ICT specially adapted for sequence analysis involving nucleotides or amino acids
    • G16B30/10—Sequence alignment; Homology search
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
    • G16B35/00—ICT specially adapted for in silico combinatorial libraries of nucleic acids, proteins or peptides
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
    • G16B35/00—ICT specially adapted for in silico combinatorial libraries of nucleic acids, proteins or peptides
    • G16B35/20—Screening of libraries
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16C—COMPUTATIONAL CHEMISTRY; CHEMOINFORMATICS; COMPUTATIONAL MATERIALS SCIENCE
    • G16C20/00—Chemoinformatics, i.e. ICT specially adapted for the handling of physicochemical or structural data of chemical particles, elements, compounds or mixtures
    • G16C20/60—In silico combinatorial chemistry
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
    • G16B20/00—ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
    • G16B20/00—ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations
    • G16B20/50—Mutagenesis

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Theoretical Computer Science (AREA)
  • Biotechnology (AREA)
  • Evolutionary Biology (AREA)
  • Medical Informatics (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Library & Information Science (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Immunology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Computing Systems (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The present invention provides a methodology for efficiently generating and screening protein libraries for optimized proteins with desirable biological functions, such as improved binding affinity towards biologically and/or therapeutically important target molecules. The process is carried out computationally in a high throughput manner by mining the ever-expanding databases of protein sequences of all organisms, especially human. In one embodiment, a method for constructing a library of designed proteins, comprising the steps of: providing an amino acid sequence derived from a lead protein, the amino acid sequence being designated as a lead sequence; comparing the lead sequence with a plurality of tester protein sequences; and selecting from the plurality of tester protein sequences at least two peptide segments that have at least 15% sequence identity with the lead sequence, the selected peptide segments forming a hit library; and forming a library of designed proteins by substituting the lead sequence with the hit library. The library of designed proteins can be expressed in vitro or in vivo to produce a library of recombinant proteins that can be screened for novel or improved function(s) over the lead protein, such as an antibody against therapeutically important target.
PCT/US2003/016037 2002-05-20 2003-05-20 Generation and selection of protein library in silico WO2003099999A2 (en)

Priority Applications (5)

Application Number Priority Date Filing Date Title
CN038173603A CN1672160B (en) 2002-05-20 2003-05-20 Method for constructing antibody library based on precursor antibody structure
AU2003248548A AU2003248548B2 (en) 2002-05-20 2003-05-20 Generation and selection of protein library in silico
JP2004508241A JP2005526518A (en) 2002-05-20 2003-05-20 Insilico creation and selection of protein libraries
CA002485732A CA2485732A1 (en) 2002-05-20 2003-05-20 Generation and selection of protein library in silico
EP03755415A EP1514216A4 (en) 2002-05-20 2003-05-20 Generation and selection of protein library in silico

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US10/153,176 US20030022240A1 (en) 2001-04-17 2002-05-20 Generation and affinity maturation of antibody library in silico
US10/153,176 2002-05-20
US10/153,159 US7117096B2 (en) 2001-04-17 2002-05-20 Structure-based selection and affinity maturation of antibody library
US10/153,159 2002-05-20

Publications (2)

Publication Number Publication Date
WO2003099999A2 WO2003099999A2 (en) 2003-12-04
WO2003099999A3 true WO2003099999A3 (en) 2004-09-16

Family

ID=29586304

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2003/016037 WO2003099999A2 (en) 2002-05-20 2003-05-20 Generation and selection of protein library in silico

Country Status (7)

Country Link
EP (1) EP1514216A4 (en)
JP (1) JP2005526518A (en)
CN (1) CN1672160B (en)
AU (1) AU2003248548B2 (en)
CA (1) CA2485732A1 (en)
SG (1) SG135053A1 (en)
WO (1) WO2003099999A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7462698B2 (en) 2005-07-22 2008-12-09 Y's Therapeutics Co., Ltd. Anti-CD26 antibodies and methods of use thereof

Families Citing this family (42)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7667004B2 (en) * 2001-04-17 2010-02-23 Abmaxis, Inc. Humanized antibodies against vascular endothelial growth factor
AU2004202566B2 (en) * 2004-06-11 2011-07-21 Li-Te Chin Method for Producing Human Antibodies with Properties of Agonist, Antagonist, or Inverse Agonist
AU2005287406B2 (en) 2004-07-26 2011-08-18 Biogen Ma Inc. Anti-CD154 antibodies
EP1824987B1 (en) 2004-11-16 2013-08-21 KaloBios Pharmaceuticals, Inc. Immunoglobulin variable region cassette exchange
EP1869084A2 (en) 2005-03-04 2007-12-26 Biogen Idec MA Inc. Methods of humanizing immunoglobulin variable regions through rational modification of complementarity determining residues
JP5374359B2 (en) * 2006-03-17 2013-12-25 バイオジェン・アイデック・エムエイ・インコーポレイテッド Stabilized polypeptide compounds
CN103541018A (en) 2006-10-02 2014-01-29 航道生物技术有限责任公司 Design and construction of diverse synthetic peptide and polypeptide librarie
EP2118795A4 (en) * 2007-01-31 2010-07-07 Sundia Meditech Company Ltd Methods, systems, algorithyms and means for describing the possible conformations of actual and theoretical proteins and for evaluating actual and theoretical proteins with respect to folding, overall shape and structural motifs
DK2356270T3 (en) * 2008-11-07 2016-12-12 Fabrus Llc Combinatorial antibody libraries and uses thereof
NZ596837A (en) * 2009-06-17 2014-02-28 Abbvie Biotherapeutics Inc Anti-vegf antibodies and their uses
JP2013508287A (en) * 2009-10-14 2013-03-07 ヤンセン バイオテツク，インコーポレーテツド Methods for affinity maturation of antibodies
WO2013171810A1 (en) 2012-05-14 2013-11-21 パナソニック株式会社 Method for obtaining heat-resistant antibody-presenting phage
US20140154255A1 (en) 2012-11-30 2014-06-05 Abbvie Biotherapeutics Inc. Anti-vegf antibodies and their uses
LT2951579T (en) * 2013-01-31 2024-05-27 Codexis, Inc. Methods, systems, and software for identifying bio-molecules using models of multiplicative form
CN104805507B (en) * 2014-01-29 2019-01-22 杭州康万达医药科技有限公司 phage display library and its application and preparation method
CA2989383A1 (en) * 2014-07-07 2016-01-14 Yeda Research And Development Co. Ltd. Method of computational protein design
GB201500464D0 (en) * 2015-01-12 2015-02-25 Crescendo Biolog Ltd Method of producing optimised therapeutic molecules
EP3262216A4 (en) * 2015-02-24 2018-12-05 Academia Sinica A phage-displayed single-chain variable fragment library
CN104789555A (en) * 2015-04-29 2015-07-22 江南大学 Method of expressing regulatory element based on evolution gene by synthesizing single-chain DNA library
JP6558754B2 (en) 2015-08-07 2019-08-14 富士通株式会社 Information processing apparatus, index dimension extraction method, and index dimension extraction program
UA125432C2 (en) 2015-09-23 2022-03-09 Дженентек, Інк. Optimized variants of anti-vegf antibodies
CN106290908A (en) * 2016-08-07 2017-01-04 查文娟 A kind of for kidney injury detection test kit
WO2018052131A1 (en) * 2016-09-16 2018-03-22 国立大学法人大阪大学 Immunological entity clustering software
CN106905435B (en) * 2017-03-13 2020-04-10 武汉海沙百得生物技术有限公司 Method for preparing binding protein based on protein A mutant
EA201992476A1 (en) * 2017-04-18 2020-02-25 Икс-Чем, Инк. METHODS FOR IDENTIFICATION OF COMPOUNDS
WO2019036055A2 (en) 2017-08-18 2019-02-21 Ignite Biosciences, Inc. Methods of selecting binding reagents
WO2019099882A2 (en) * 2017-11-20 2019-05-23 Nantbio, Inc. mRNA DISPLAY ANTIBODY LIBRARY AND METHODS
CN108304691B (en) * 2018-02-09 2021-05-18 北京矿冶科技集团有限公司 Fragment-based flotation agent molecule design method
CN108491692B (en) * 2018-03-09 2023-07-21 中国科学院生态环境研究中心 A method for constructing antibiotic resistance gene database
CN108763870B (en) * 2018-05-09 2021-08-03 浙江工业大学 A construction method of multi-domain protein Linker
CN109002690B (en) * 2018-06-08 2021-05-18 济南大学 Method for predicting side chain structure of mutant amino acid by constructing CHARMM ROTAMERS force field
CN108959846B (en) * 2018-07-03 2021-09-14 南昌立德生物技术有限公司 Affinity free energy decomposition algorithm for computer-aided pilot medicament optimization design
CN109243526B (en) * 2018-07-12 2021-08-03 浙江工业大学 A protein structure prediction method based on the intersection of specific fragments
CN109086568B (en) * 2018-08-16 2022-03-11 福建工程学院 Computer antibody combination mutation evolution system and method, information data processing terminal
CN109801679B (en) * 2019-01-15 2021-02-02 广州柿宝生物科技有限公司 A Mathematical Sequence Reconstruction Method for Long Chain Molecules
WO2020208555A1 (en) * 2019-04-09 2020-10-15 Eth Zurich Systems and methods to classify antibodies
WO2020246617A1 (en) * 2019-06-07 2020-12-10 中外製薬株式会社 Information processing system, information processing method, program, and method for producing antigen-binding molecule or protein.
AU2021210992A1 (en) * 2020-01-23 2022-08-04 Massachusetts Institute Of Technology Methods of functionally screening biological sequence fragments
JP2023513314A (en) * 2020-02-13 2023-03-30 ザイマージェン インコーポレイテッド Metagenome library and natural product discovery platform
CN111370074B (en) * 2020-02-27 2023-07-07 北京晶泰科技有限公司 Method and device for generating molecular sequence and computing equipment
CN111462815B (en) * 2020-03-27 2023-05-02 上海祥耀生物科技有限责任公司 Construction method and device of antibody library
CN112102883B (en) * 2020-08-20 2023-12-08 深圳华大生命科学研究院 Base sequence coding method and system in FASTQ file compression

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU3415395A (en) * 1994-08-26 1996-03-22 Eli Lilly And Company Antibody constructs with cdr switched variable regions
WO2002084277A1 (en) * 2001-04-17 2002-10-24 Abmaxis, Inc. Structure-based construction of human antibody library

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHOTHIA ET AL: "Structural determinants in the sequences of immunoglobulin variable domain", JOURNAL OF MOLECULAR BIOLOGY, vol. 278, 1998, pages 457 - 479, XP004453679 *
KNAPPIK ET AL: "Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides", JOURNAL OF MOLECULAR BIOLOGY, vol. 296, 2000, pages 57 - 86, XP004461525 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7462698B2 (en) 2005-07-22 2008-12-09 Y's Therapeutics Co., Ltd. Anti-CD26 antibodies and methods of use thereof

Also Published As

Publication number Publication date
CN1672160A (en) 2005-09-21
CA2485732A1 (en) 2003-12-04
WO2003099999A2 (en) 2003-12-04
AU2003248548B2 (en) 2010-03-11
EP1514216A2 (en) 2005-03-16
SG135053A1 (en) 2007-09-28
AU2003248548A1 (en) 2003-12-12
EP1514216A4 (en) 2010-01-06
CN1672160B (en) 2010-06-09
JP2005526518A (en) 2005-09-08

Similar Documents

Publication Publication Date Title
WO2003099999A3 (en) Generation and selection of protein library in silico
HK1072957A1 (en) In vitro peptide expression libraray
JP4972191B2 (en) Methods and compositions for peptide, protein and peptide mimetic synthesis
ATE297462T1 (en) LIBRARY FOR IN VITRO EXPRESSION OF PEPTIDES OR PROTEINS
WO2002029032A3 (en) Whole cell engineering by mutagenizing a substantial portion of a starting genome, combining mutations, and optionally repeating
EP1774019A4 (en) Look-through mutagenesis for developing altered polypeptides with enhanced properties
WO2005003345A3 (en) Look-through mutagenesis
JP2007127631A (en) A method for selective separation of multiply charged peptides applicable to quantitative proteomics
DE69827810D1 (en) METHOD FOR THROWING BANKS BY CHIMERAL BINDING PETIDES
Biemann Laying the groundwork for proteomics: mass spectrometry from 1958 to 1988
EA200100160A1 (en) NEW METHODS OF IDENTIFICATION OF BIOMOLECULES OF LIGANDS AND TARGETS
CA2345287A1 (en) Tgf-beta superfamily mutant members, including morphogenic proteins
JP2004531225A5 (en)
Ishida et al. Engineering ribosomal machinery for noncanonical amino acid incorporation
AU8649698A (en) Cell cycle-regulating proteins
Cutfield et al. The amino‐acid sequences of sculpin islet somatostatin‐28 and peptide YY
Katoh et al. Advances in in vitro genetic code reprogramming in 2014–2017
Moss et al. Solid phase peptide synthesis on JandaJelTM resin
WO2003073351A3 (en) Screening process
EP2354156A3 (en) Nucleic acids and proteins of insect OR83B odorant receptor genes and uses thereof
AU2003202689A8 (en) Methods of screening molecular libraries for active molecules that restore the function of target proteins and peptides that restore the function of mutant p53
Spirin Testing the classical two-tRNA-site model for the ribosomal elongation cycle
Thapa et al. t‐boc synthesis of huwentoxin‐i through native chemical ligation incorporating a trifluoromethanesulfonic acid cleavage strategy
JP7654312B2 (en) Method for screening sequences that modulate the display efficiency of polynucleotide display methods
Schiff Establishing Peptide-AS-MS as a novel high-throughput screening technology to provide a pipeline for discovery of new affinity reagents in New Zealand

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NI NO NZ OM PH PL PT RO RU SC SD SE SG SK SL TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2485732

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2003248548

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 2004508241

Country of ref document: JP

WWE Wipo information: entry into national phase

Ref document number: 2003755415

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 20038173603

Country of ref document: CN

WWP Wipo information: published in national office

Ref document number: 2003755415

Country of ref document: EP

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载