Isolating recombinant antibodies against specific protein morphologies using atomic force microscopy and phage display technologies

Hedieh Barkhordarian, Sharareh Emadi, Philip Schulz, Michael Sierks

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Isolation of antibodies to antigens that are either unstable, exist in multiple morphologies or have very limited availability can be prohibitively difficult. Here we describe a novel technique combining the capabilities of phage display antibody technology and atomic force microscopy (AFM) that is used to isolate antibody fragments that bind to a specific morphology of the target antigen, α-synuclein. AFM imaging allows us to both visualize the presence and morphology of the target antigen as well as to monitor the efficiency of each step in the bio-panning process. We demonstrate that phage displayed antibodies specific to the target antigen morphology can be isolated after only two rounds of selection. The target antigen, α-synuclein, has been correlated with the Parkinson's disease (PD). Accumulation of α-synuclein fibrillar aggregates into Lewy body inclusions is a hallmark feature of PD. While α-synuclein can form several different aggregate morphologies including oligomers, protofibrils and fibrils, the role of these morphologies in the progression of PD is not known. The successful selection of the recombinant antibody described here can have potential therapeutic value since the single-chain fragment variable (scFv) can be expressed intracellularly to control folding and toxicity of the specific protein aggregates.

Original languageEnglish (US)
Pages (from-to)497-502
Number of pages6
JournalProtein Engineering, Design and Selection
Volume19
Issue number11
DOIs
StatePublished - Nov 2006

Fingerprint

Synucleins
Bacteriophages
Atomic Force Microscopy
Antibodies
Antigens
Atomic force microscopy
Display devices
Technology
Proteins
Parkinson Disease
Lewy Bodies
Single-Chain Antibodies
Immunoglobulin Fragments
Oligomers
Toxicity
Availability
Imaging techniques

Keywords

  • Antibodies
  • Atomic force microscopy
  • Bio-panning
  • phage display
  • Protein morphology

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology

Cite this

Isolating recombinant antibodies against specific protein morphologies using atomic force microscopy and phage display technologies. / Barkhordarian, Hedieh; Emadi, Sharareh; Schulz, Philip; Sierks, Michael.

In: Protein Engineering, Design and Selection, Vol. 19, No. 11, 11.2006, p. 497-502.

Research output: Contribution to journalArticle

@article{e5aec3a11b1148b9835239aac0de3b05,
title = "Isolating recombinant antibodies against specific protein morphologies using atomic force microscopy and phage display technologies",
abstract = "Isolation of antibodies to antigens that are either unstable, exist in multiple morphologies or have very limited availability can be prohibitively difficult. Here we describe a novel technique combining the capabilities of phage display antibody technology and atomic force microscopy (AFM) that is used to isolate antibody fragments that bind to a specific morphology of the target antigen, α-synuclein. AFM imaging allows us to both visualize the presence and morphology of the target antigen as well as to monitor the efficiency of each step in the bio-panning process. We demonstrate that phage displayed antibodies specific to the target antigen morphology can be isolated after only two rounds of selection. The target antigen, α-synuclein, has been correlated with the Parkinson's disease (PD). Accumulation of α-synuclein fibrillar aggregates into Lewy body inclusions is a hallmark feature of PD. While α-synuclein can form several different aggregate morphologies including oligomers, protofibrils and fibrils, the role of these morphologies in the progression of PD is not known. The successful selection of the recombinant antibody described here can have potential therapeutic value since the single-chain fragment variable (scFv) can be expressed intracellularly to control folding and toxicity of the specific protein aggregates.",
keywords = "Antibodies, Atomic force microscopy, Bio-panning, phage display, Protein morphology",
author = "Hedieh Barkhordarian and Sharareh Emadi and Philip Schulz and Michael Sierks",
year = "2006",
month = "11",
doi = "10.1093/protein/gzl036",
language = "English (US)",
volume = "19",
pages = "497--502",
journal = "Protein Engineering, Design and Selection",
issn = "1741-0126",
publisher = "Oxford University Press",
number = "11",

}

TY - JOUR

T1 - Isolating recombinant antibodies against specific protein morphologies using atomic force microscopy and phage display technologies

AU - Barkhordarian, Hedieh

AU - Emadi, Sharareh

AU - Schulz, Philip

AU - Sierks, Michael

PY - 2006/11

Y1 - 2006/11

N2 - Isolation of antibodies to antigens that are either unstable, exist in multiple morphologies or have very limited availability can be prohibitively difficult. Here we describe a novel technique combining the capabilities of phage display antibody technology and atomic force microscopy (AFM) that is used to isolate antibody fragments that bind to a specific morphology of the target antigen, α-synuclein. AFM imaging allows us to both visualize the presence and morphology of the target antigen as well as to monitor the efficiency of each step in the bio-panning process. We demonstrate that phage displayed antibodies specific to the target antigen morphology can be isolated after only two rounds of selection. The target antigen, α-synuclein, has been correlated with the Parkinson's disease (PD). Accumulation of α-synuclein fibrillar aggregates into Lewy body inclusions is a hallmark feature of PD. While α-synuclein can form several different aggregate morphologies including oligomers, protofibrils and fibrils, the role of these morphologies in the progression of PD is not known. The successful selection of the recombinant antibody described here can have potential therapeutic value since the single-chain fragment variable (scFv) can be expressed intracellularly to control folding and toxicity of the specific protein aggregates.

AB - Isolation of antibodies to antigens that are either unstable, exist in multiple morphologies or have very limited availability can be prohibitively difficult. Here we describe a novel technique combining the capabilities of phage display antibody technology and atomic force microscopy (AFM) that is used to isolate antibody fragments that bind to a specific morphology of the target antigen, α-synuclein. AFM imaging allows us to both visualize the presence and morphology of the target antigen as well as to monitor the efficiency of each step in the bio-panning process. We demonstrate that phage displayed antibodies specific to the target antigen morphology can be isolated after only two rounds of selection. The target antigen, α-synuclein, has been correlated with the Parkinson's disease (PD). Accumulation of α-synuclein fibrillar aggregates into Lewy body inclusions is a hallmark feature of PD. While α-synuclein can form several different aggregate morphologies including oligomers, protofibrils and fibrils, the role of these morphologies in the progression of PD is not known. The successful selection of the recombinant antibody described here can have potential therapeutic value since the single-chain fragment variable (scFv) can be expressed intracellularly to control folding and toxicity of the specific protein aggregates.

KW - Antibodies

KW - Atomic force microscopy

KW - Bio-panning

KW - phage display

KW - Protein morphology

UR - http://www.scopus.com/inward/record.url?scp=33750291199&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33750291199&partnerID=8YFLogxK

U2 - 10.1093/protein/gzl036

DO - 10.1093/protein/gzl036

M3 - Article

C2 - 16984950

AN - SCOPUS:33750291199

VL - 19

SP - 497

EP - 502

JO - Protein Engineering, Design and Selection

JF - Protein Engineering, Design and Selection

SN - 1741-0126

IS - 11

ER -