Advanced

Recombinant Antibodies – Tools for building proteomic platforms

Säll, Anna LU (2015)
Abstract
This thesis, which is based on four original papers, covers different aspects of the evolvement and development of proteomic platforms and the construction of human synthetic antibody libraries for the isolation of antibody reagents has been very central. There is an accelerating need for technologies capable of providing proteomic analysis of high sensitivity with good accuracy and an overall broad coverage as well as for biomarker validation. Paper I, focused on the design and construction of human synthetic antibody libraries for the isolation of scFv antibodies suitable for the use in different affinity proteomic platforms. High-throughput procedures for the isolation as well as screening of antibodies was established and provides a... (More)
This thesis, which is based on four original papers, covers different aspects of the evolvement and development of proteomic platforms and the construction of human synthetic antibody libraries for the isolation of antibody reagents has been very central. There is an accelerating need for technologies capable of providing proteomic analysis of high sensitivity with good accuracy and an overall broad coverage as well as for biomarker validation. Paper I, focused on the design and construction of human synthetic antibody libraries for the isolation of scFv antibodies suitable for the use in different affinity proteomic platforms. High-throughput procedures for the isolation as well as screening of antibodies was established and provides a pipeline capable of selecting antibodies against a large number of target antigens in parallel. Furthermore, scFv antibodies generated from these antibody libraries were validated for the use in antibody microarrays and were also utilized in various affinity-based proteomic platforms. In paper II and III, two different approaches are taken to address problems associated with proteomic analysis. Technologies capable of biomarker validation are highly required for increasing the implementation of protein biomarkers in clinical use. A method combining affinity enrichment with the use of scFv antibodies and mass spectrometric read-out were developed in paper II. The platform allows multiplexed detection of low abundant target proteins in human serum samples. Paper III focused on establishing a platform to facilitate the detection and investigation of bacterial virulence factors by using Streptococcus pyogenes as a model system. Bacterial infections are still a serious health problem around the world and analysis exploring bacterial strategies promoting survival and proliferation within the human host are of great importance. The performance of an affinity-based platform is dependent on the functionality of the used affinity reagents. In paper IV, a semi-targeted platform for biomarker discovery were enhanced by taking advantage of an oriented antibody immobilization strategy. In conclusion, this thesis presents a pipeline for development of human scFv antibodies suitable for proteomic analysis as well as the establishment and evolvement of various proteomic platforms. (Less)
Please use this url to cite or link to this publication:
author
supervisor
opponent
  • Persson, Mats, Karolinska Institutet
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Antibody development, recombinant scFv antibodies, phage display, affinity proteomics, mass spectrometry
defense location
Lecture hall, Medicon Village, Lund University, Lund
defense date
2015-12-18 09:15
ISBN
978-91-7623-569-0
language
English
LU publication?
yes
id
735e66ff-633c-4b6e-8466-c4f7b0d20c50 (old id 8228774)
date added to LUP
2015-11-24 07:24:13
date last changed
2016-09-19 08:45:16
@misc{735e66ff-633c-4b6e-8466-c4f7b0d20c50,
  abstract     = {This thesis, which is based on four original papers, covers different aspects of the evolvement and development of proteomic platforms and the construction of human synthetic antibody libraries for the isolation of antibody reagents has been very central. There is an accelerating need for technologies capable of providing proteomic analysis of high sensitivity with good accuracy and an overall broad coverage as well as for biomarker validation. Paper I, focused on the design and construction of human synthetic antibody libraries for the isolation of scFv antibodies suitable for the use in different affinity proteomic platforms. High-throughput procedures for the isolation as well as screening of antibodies was established and provides a pipeline capable of selecting antibodies against a large number of target antigens in parallel. Furthermore, scFv antibodies generated from these antibody libraries were validated for the use in antibody microarrays and were also utilized in various affinity-based proteomic platforms. In paper II and III, two different approaches are taken to address problems associated with proteomic analysis. Technologies capable of biomarker validation are highly required for increasing the implementation of protein biomarkers in clinical use. A method combining affinity enrichment with the use of scFv antibodies and mass spectrometric read-out were developed in paper II. The platform allows multiplexed detection of low abundant target proteins in human serum samples. Paper III focused on establishing a platform to facilitate the detection and investigation of bacterial virulence factors by using Streptococcus pyogenes as a model system. Bacterial infections are still a serious health problem around the world and analysis exploring bacterial strategies promoting survival and proliferation within the human host are of great importance. The performance of an affinity-based platform is dependent on the functionality of the used affinity reagents. In paper IV, a semi-targeted platform for biomarker discovery were enhanced by taking advantage of an oriented antibody immobilization strategy. In conclusion, this thesis presents a pipeline for development of human scFv antibodies suitable for proteomic analysis as well as the establishment and evolvement of various proteomic platforms.},
  author       = {Säll, Anna},
  isbn         = {978-91-7623-569-0},
  keyword      = {Antibody development,recombinant scFv antibodies,phage display,affinity proteomics,mass spectrometry},
  language     = {eng},
  title        = {Recombinant Antibodies – Tools for building proteomic platforms},
  year         = {2015},
}