Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Aachen
Apprimus Verlag
2017
|
| Ausgabe: | 1. Auflage |
| Schriftenreihe: | Ergebnisse aus den Naturwissenschaften
11 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | II, 97, XVIII Seiten Illustrationen, Diagramme 21 cm |
| ISBN: | 9783863595142 3863595149 |
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| 264 | 1 | |a Aachen |b Apprimus Verlag |c 2017 | |
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| 653 | |a Naturwissenschaften | ||
| 653 | |a Plasmodium falciparum | ||
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Datensatz im Suchindex
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|---|---|
| adam_text | TABLE OF CONTENTS
1
INTRODUCTION.........................................................................................................................
1
1.1 M
ALARIA...........................................................................................................................................................1
1.2 LIFE CYCLE OF PLASMODIUM FALCIPARUM
....................
1.3 VACCINES AGAINST MALARIA
.........................................
1.3.1 PRE-ERYTHROCYTIC VACCIN ES
..............................
1.3.2 BLOOD STAGE VACCINES.......................................
1.3.3 TRANSMISSION-BLOCKING VACCINES
...................
1.3.3.1 THE TB V CANDIDATE PFS48/45
....................
1.3.3.2 THE TB V CANDIDATE PFS230
.......................
1.3.3.3 THE TBV CANDIDATE P /G A P 50
...................
1.3.3.4 SEXUAL STAGE FUSION VACCINE CANDIDATES
1.3.3.5 TB V CANDIDATES IN (PRE-) CLINICAL
TRIALS....................................................................................
10
1.4 PLANTS AS PRODUCTION
PLATFORMS................................................................................................................10
1.5 OBJECTIVES OF THE
RESEARCH........................................................................................................................11
2 MATERIALS AND
METHODS......................................................................................................15
2.1 CHEMICALS AND
CONSUMABLES..................................................................................................................
15
2.2 ENZYMES, ANTIBODIES AND REACTION KITS
................................................................................................
15
2.3 MEDIA, SOLUTION AND
BUFFERS....................................................................................................................
16
2.4
VECTORS..........................................................................................................................................................
IS
2.5 GENE
VARIANTS.............................................................................................................................................20
2.6 SYNTHETIC
OLIGONUCLEOTIDES.......................................................................................................................20
2.7 ORGANISM
S..................................................................................................................................................
22
2.8 MOLECULAR BIOLOGICAL M
ETHODS................................................................................................................
22
2.8.1 CULTIVATION OF BACTERIA AND PREPARATION OF GLYCEROLSTOCKS
......................................................
22
2.8.2 PREPARATION OF CHEMICAL OR ELECTROCOMPATENT
CELLS..................................................................22
2.8.3 HEAT TRANSFORMATION OF E. COLI AND ELECTROPORATION OF A.
TUMEFACIENS................................23
2.8.4 PLASMID ISOLATION FROM E. COLI AND RESTRICTION OF D M A
............................................................. 23
2.8.5 RNA ISOLATION, CDNA AMPLIFICATION AND LINKER
ATTACHMENT.....................................................23
2.8.6 AGAROSE GEL ELECTROPHORESIS AND ISOLATION OF
DNA-FRAGMENTS..............................................24
2.8.7
LIGATION...............................................................................................................................................
24
2.8.8 POLYMERASE CHAIN REACTION (PCR), 5*RACE PCR AND SANGER SEQUENCING
.....................
24
2.9 PROTEIN CHEMICAL AND IMMUNOLOGICAL M
ETHODS..................................................................................
26
2.9.1 PROTEIN EXTRACTION AND
PURIFICATION...............................................................................................
26
2.9.2 IMMUNOAFFINITY CHROMATOGRAPHY (IA C
)........................................................................................
26
2.9.3 SDS-PAG E AND IMMUNOBLOT
ANALYSIS........................................................................................27
2.9.4 ENZYME-LINKED IMMUNOSORBENT
ASSAY........................................................................................27
2.9.5 SURFACE PLASMON RESONANCE
MEASUREMENT...............................................................................28
2.9.6 BRADFORD AND BCA
ASSAY...............................................................................................................
28
2.9.7 PROTEIN QUANTIFICATION USING FLUORESCENCE MEASUREMENT
.......................................................
29
2.10
PLANTS............................................................................................................................................................29
2.10.1 CULTIVATION OF NICOTIANA
BENTHAMIANA.........................................................................................
29
2.10.2 TRANSIENT PROTEIN EXPRESSION IN N. BENTHAMIANA
.....................................................................
29
2.11 EXPERIMENTS WITH HUMAN PLASMA, ANIMALS AND PLASMODIUM FALCIPARUM
......................................29
2.11.1 PLASMA REACTIVITIES OF AFRICAN SERUM SAM
PLES..........................................................................29
2.11.2 IMMUNIZATION OF MICE AND RABBITS AND DETERMINATION OF
TITERS..............................................30
2.11.3 GENERATION OF HYBRIDOMA CELL LINES AND HYBRIDOMA
RESCUE...................................................30
2.11.4 ASSAYS WITH P.
FALCIPAMM..............................................................................................................
31
3
RESULTS................................................................................................................................
33
3.1
P/S48/45.......................................................................................................................................................
33
3.1.1 CLONING OF EXPRESSION VECTORS FOR
PFS48/45...............................................................................33
3.1.2 EXPRESSION OF PFS48/45 GLYCO-KNOCKOUT VARIANTS
.....................................................................
36
3.1.3 EXPRESSION OF GLYCOSYLATED PFS48/45
VARIANTS............................................................................36
3.2
PFS230...........................................................................................................................................................41
3.2.1 CLONING OF EXPRESSION VECTORS FOR P
FS230..................................................................................
41
3.2.2 TRANSIENT EXPRESSION IN
PLANTS......................................................................................................42
3.2.3 DETERMINATION OF ANTIGEN-SPECIFIC TITERS AGAINST P /S230_C 0
.................................................
43
3.2.4 IMMUNOFLUORESCENCE
ASSAY...........................................................................................................44
3.3 P /G A P
50......................................................................................................................................................
46
D (O CO N O) O L O L A IS)
3.3.1 CLONING OF EXPRESSION VECTORS FOR
P/GAPSO...............................................................................46
3.3.2 EXPRESSION IN PLANTS AND
PURIFICATION...........................................................................................47
3.3.3 DETERMINATION OF ANTIGEN-SPECIFIC TITERS IN
RABBITS....................................................................
49
3.3.4 IMMUNOFLUORESCENCE
ASSAYS.........................................................................................................49
3.3.5 ZYGOTE INHIBITION ASSAY (Z IA
).........................................................................................................
50
3.3.6 TRANSMISSION-BLOCKING ASSAY (T B A
)............................................................................................
51
3.4 SEXUAL STAGE FUSION
PROTEINS..................................................................................................................
53
3.4.1 CLONING OF SEXUAL STAGE FUSION CONSTRUCTS
.................................................................................
53
3.4.2 EXPRESSION IN PLANTS AND
PURIFICATION...........................................................................................55
3.4.3 DETERMINATION OF ANTIGEN-SPECIFIC
TITERS......................................................................................
57
3.4.4 PLASMA REACTIVITY FROM AFRICAN SERUM
SAMPLES.........................................................................
58
3.4.5 IMMUNOFLUORESCENCE
ASSAYS.........................................................................................................
59
3.4.6 T RANSMISSION-BLOCKING
ASSAY.........................................................................................................60
3.5 ANTIBODIES AGAINST SINGLE SEXUAL STAGE
PROTEINS................................................................................
62
3.5.1 THE PFS25-SPECIFIC MOUSE MONOCLONAL ANTIBODY 4 B 7
.............................................................
62
3.5.1.1 CLONING SEQUENCES ENCODING THE MOUSE MONOCLONAL ANTIBODY 4 B 7
...............................63
3.5.1.2 EXPRESSION IN PLANTS AND
PURIFICATION......................................................................................
64
3.5.1.3 IMMUNOAFFINITY CHROMATOGRAPHY WITH PLANT-DERIVED M
AB4B7............................................65
3.5.2 GENERATION OF MONOCLONAL ANTIBODIES AGAINST PFS230, PFS25 AND PIC
E LT O S
...................
66
3.5.3 HYBRIDOMA RESCUE OF ANTIBODIES AGAINST PFS230, PFS25 AND P /C E LT
O S
...........................
67
4 DISCUSSION AND
CONCLUSION..............................................................................................
69
4.1 EVALUATION OF P
/S48/45...............................................................................................................................69
4.2 EVALUATION OF P
/S230..................................................................................................................................71
4.3 EVALUATION OF
PK3A P
50.............................................................................................................................
72
4.4 EVALUATION OF SEXUAL STAGE FUSION
PROTEINS..........................................................................................74
4.5 EVALUATION OF ANTIBODIES AGAINST SINGLE SEXUAL STAGE
ANTIGENS..................................................... 76
4.6 GENERAL CONCLUSION AND
OUTLOOK............................................................................................................
76
5
SUMMARY.............................................................................................................................
83
6 DEUTSCHE
ZUSAMMENFASSUNG..........................................................................................85
7
REFERENCES..........................................................................................................................87
8
APPENDIX................................................................................................................................
I
8.1 REGISTER OF
FIGURES..........................................................................................................................................
I
8.2 REGISTER OF
TABLES..........................................................................................................................................
II
6.3
ABBREVIATIONS................................................................................................................................................ILL
6.4 SUPPLEMENTAL
FIGURES.................................................................................................................................
VI
CURRICULUM
VITAE......................................................................................................................XVII
|
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| dewey-ones | 500 - Natural sciences and mathematics |
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| dewey-search | 500 |
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| discipline | Allgemeine Naturwissenschaft |
| edition | 1. Auflage |
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| indexdate | 2024-07-10T07:52:29Z |
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| language | English |
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| physical | II, 97, XVIII Seiten Illustrationen, Diagramme 21 cm |
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| spelling | Beiß, Véronique Verfasser (DE-588)1136736816 aut Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum vorgelegt von Véronique Beiß 1. Auflage Aachen Apprimus Verlag 2017 II, 97, XVIII Seiten Illustrationen, Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Ergebnisse aus den Naturwissenschaften 11 Dissertation Rheinisch-Westfälische Technische Hochschule Aachen 2017 Impfstoff (DE-588)4026655-2 gnd rswk-swf Malaria (DE-588)4037197-9 gnd rswk-swf Plasmodium falciparum (DE-588)4250521-5 gnd rswk-swf Antigen (DE-588)4002272-9 gnd rswk-swf Fachhochschul-/Hochschulausbildung Erwachsenenbildung Biologie Naturwissenschaften Plasmodium falciparum (DE-588)4113937-9 Hochschulschrift gnd-content Malaria (DE-588)4037197-9 s Impfstoff (DE-588)4026655-2 s Plasmodium falciparum (DE-588)4250521-5 s Antigen (DE-588)4002272-9 s DE-604 Apprimus Verlag (DE-588)1068101474 pbl Ergebnisse aus den Naturwissenschaften 11 (DE-604)BV041770386 11 B:DE-101 application/pdf http://d-nb.info/1135291349/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029822031&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Beiß, Véronique Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum Ergebnisse aus den Naturwissenschaften Impfstoff (DE-588)4026655-2 gnd Malaria (DE-588)4037197-9 gnd Plasmodium falciparum (DE-588)4250521-5 gnd Antigen (DE-588)4002272-9 gnd |
| subject_GND | (DE-588)4026655-2 (DE-588)4037197-9 (DE-588)4250521-5 (DE-588)4002272-9 (DE-588)4113937-9 |
| title | Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum |
| title_auth | Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum |
| title_exact_search | Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum |
| title_full | Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum vorgelegt von Véronique Beiß |
| title_fullStr | Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum vorgelegt von Véronique Beiß |
| title_full_unstemmed | Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum vorgelegt von Véronique Beiß |
| title_short | Multi-target malaria vaccines against the sexual stage of Plasmodium falciparum |
| title_sort | multi target malaria vaccines against the sexual stage of plasmodium falciparum |
| topic | Impfstoff (DE-588)4026655-2 gnd Malaria (DE-588)4037197-9 gnd Plasmodium falciparum (DE-588)4250521-5 gnd Antigen (DE-588)4002272-9 gnd |
| topic_facet | Impfstoff Malaria Plasmodium falciparum Antigen Hochschulschrift |
| url | http://d-nb.info/1135291349/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029822031&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV041770386 |
| work_keys_str_mv | AT beißveronique multitargetmalariavaccinesagainstthesexualstageofplasmodiumfalciparum AT apprimusverlag multitargetmalariavaccinesagainstthesexualstageofplasmodiumfalciparum |
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