Holdings: Generation and characterization of novel antibodies to M. tuberculosis antigens

Generation and characterization of novel antibodies to M. tuberculosis antigens:

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Bibliographic Details
Main Author:	Gautam, Kritan 1986- (Author)
Format:	Thesis Book
Language:	English
Published:	Jena April 2018
Subjects:	Hochschulschrift
Online Access:	Inhaltsverzeichnis Inhaltsverzeichnis
Physical Description:	118, xxxiv Seiten Illustrationen 29 cm

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MARC


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adam_text	TABLE OF CONTENTS TABLE OF CONTENTS....................................................................................................................I UNITS LIST................................................................................................................................. IV ABBREVIATIONS LIST....................................................................................................................V SUMMARY................................................................................................................................. 1 ZUSAMMENFASSUNG................................................................................................................ 2 1 INTRODUCTION...................... 3 1.1 ANTIBODIES............................................. 3 1.1.1 BIOLOGY AND STRUCTURE OF ANTIBODIES.............................................................. .. 3 1.1.2 EXPERIMENTAL GENERATION OF ANTIBODIES............................................................4 1.1.3 DISPLAY TECHNIQUE.............................................................................................. 8 1.1.4 RECOMBINANT ANTIBODIES................................................................................... 9 1.2 TUBERCULOSIS (T B )................................................................................................... 10 1.2.1 BACKGROUND....................................................................................................... 10 1.2.2 TB PATHOGENESIS .............................................................................................. 11 1.2.3 DIAGNOSIS OF TUBERCULOSIS: STATE OF THE ART......................................................12 1.2.4 TUBERCULOSIS ANTIGENS WITH DIAGNOSTIC POTENTIAL............................................13 2 AIM OF THE W ORK............................................................................................................17 3 MATERIALS AND METHODS...............................................................................................18 3.1 MATERIALS................................................................................................................... 18 3.1.1 LABORATORY EQUIPMENT......................................................................................18 3.1.2 LAB SUPPLIES AND CONSUMABLES...................................................................... 19 3.1.3 CHEMICALS, BUFFERS AND SOLUTIONS...................................................................20 3.1.4 CELL CULTURE MEDIUM.........................................................................................26 3.1.5 ENZYMES AND MOLECULAR WEIGHT MARKERS ..................................................... 27 3.1.6 ANTIBODIES......................................................................................................... 28 3.1.7 PLASMIDS 29 3.1.8 BACTERIAL STRAINS, YEAST STRAIN, AND CELL LINES .................................................. 30 3.1.9 SOFTWARES............................................................................................................... 31 3.1.10 ANTIBIOTICS.............................................................................................................. 31 3.1.11DNA PRIMERS..........................................................................................................32 3.1.12 LABORATORY KITS...................................................................................................... 35 3.2 METHODS......................................................................................................................... 35 3.2.1 MOLECULAR BIOLOGICAL METHODS.............................................................................35 3.2.2 CELL CULTURE METHODS............................................................................................42 3.2.3 PROTEIN BIOCHEMICAL METHODS.............................................................................45 3.2.4 EXPERIMENTAL GENERATION OF ANTIBODIES ............................................................ 53 3.2.5 FLOW CYTOMETRY AND F A C S ................................................................................. 57 3.2.6 DETERMINATION OF K D VALUES OF ANTIBODIES........................................................58 3.2.7 ANALYSIS OF CDR REGIONS IN ANTIBODIES ............................................................ 58 3.2.8 GENERATION OF PROTEIN MODELS.............................................................................59 3.2.9 ANALYSIS OF IMMUNOGENIC EPITOPES ON ANTIGENS ............................................ 59 4 RESULTS................................................................................................................. 60 4.1 CLONING, EXPRESSION, AND PURIFICATION OF MYCOBACTERIUM TUBERCULOSIS ANTIGENS........................................................................................................................... 60 4.1.1 AMPLIFICATION OF ANTIGEN-ENCODING CDNA SEQUENCES FROM GENOMIC DNA OF M. TUBERCULOSIS................................................................................................. 60 4.1.2 GENERATION OF EXPRESSION CONSTRUCTS ENCODING MYCOBACTERIAL ANTIGENS . 64 4.1.3 EXPRESSION AND PURIFICATION OF RECOMBINANT MYCOBACTERIAL ANTIGENS AS FUSIONS WITH M B P ................................................................................................. 65 4.2 EXPERIMENTAL GENERATION OF ANTIBODIES.....................................................................72 4.2.1 GENERATION OF MONOCLONAL ANTIBODIES TO MYCOBACTERIAL ANTIGENS BY THE HYBRIDOMA TECHNOLOGY AND THEIR CHARACTERIZATION..........................................72 4.2.2 SELECTION OF ANTIBODIES FROM MOLECULAR REPERTOIRES ...................................... 82 4.2.3 GENERATION OF RECOMBINANT ANTIBODIES BINDING TO ANTIGEN (M) AG85B ..... 97 5 DISCUSSION.........................................................................................................107 6 CONCLUSION......................................................................................................... 118 REFERENCES.................................................................................................................................. VII APPENDIX......................................................................................................................................XX IN
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spelling	Gautam, Kritan 1986- Verfasser (DE-588)1179790014 aut Generation and characterization of novel antibodies to M. tuberculosis antigens by M.Sc. Kritan Gautam Jena April 2018 118, xxxiv Seiten Illustrationen 29 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Friedrich Schiller University of Jena 2018 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1176169947/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031610061&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Gautam, Kritan 1986- Generation and characterization of novel antibodies to M. tuberculosis antigens
subject_GND	(DE-588)4113937-9
title	Generation and characterization of novel antibodies to M. tuberculosis antigens
title_auth	Generation and characterization of novel antibodies to M. tuberculosis antigens
title_exact_search	Generation and characterization of novel antibodies to M. tuberculosis antigens
title_full	Generation and characterization of novel antibodies to M. tuberculosis antigens by M.Sc. Kritan Gautam
title_fullStr	Generation and characterization of novel antibodies to M. tuberculosis antigens by M.Sc. Kritan Gautam
title_full_unstemmed	Generation and characterization of novel antibodies to M. tuberculosis antigens by M.Sc. Kritan Gautam
title_short	Generation and characterization of novel antibodies to M. tuberculosis antigens
title_sort	generation and characterization of novel antibodies to m tuberculosis antigens
topic_facet	Hochschulschrift
url	http://d-nb.info/1176169947/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031610061&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
work_keys_str_mv	AT gautamkritan generationandcharacterizationofnovelantibodiestomtuberculosisantigens

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