A dimerized single-chain variable fragment system for the assessment of neutralizing activity of phage display-selected antibody fragments specific for cytomegalovirus.
(2012) In Journal of Immunological Methods 376(Online 01 Dec 2011). p.69-78- Abstract
- Abstract in Undetermined
Cytomegalovirus (CMV) causes severe sequelae in congenitally infected newborns and may cause life-threatening disease in immuno-deficient patients. Recent findings demonstrate the possibility to alleviate the disease by infusing intravenous immunoglobulin G (IgG) preparations, indicating that antibodies are an effective therapeutic option. Modern molecular methodologies, like phage display, allow for the development of specific antibodies targeting virtually any antigen, including those of CMV. However, such methodologies do not in general result in products that by themselves mediate biological activity. To facilitate a semi-high-throughput approach for functional screening in future efforts to develop... (More) - Abstract in Undetermined
Cytomegalovirus (CMV) causes severe sequelae in congenitally infected newborns and may cause life-threatening disease in immuno-deficient patients. Recent findings demonstrate the possibility to alleviate the disease by infusing intravenous immunoglobulin G (IgG) preparations, indicating that antibodies are an effective therapeutic option. Modern molecular methodologies, like phage display, allow for the development of specific antibodies targeting virtually any antigen, including those of CMV. However, such methodologies do not in general result in products that by themselves mediate biological activity. To facilitate a semi-high-throughput approach for functional screening in future efforts to develop efficacious antibodies against CMV, we have integrated two different approaches to circumvent potential bottlenecks in such efforts. Firstly, we explored an approach that permits easy transfer of antibody fragment encoding genes from commonly used phage display vectors into vectors for the production of divalent immunoglobulins. Secondly, we demonstrate that such proteins can be applied in a novel reporter-based neutralization assay to establish a proof-of-concept workflow for the generation of neutralizing antibodies against CMV. We validated our approach by showing that divalent antibodies raised against the antigenic domain (AD)-2 region of gB effectively neutralized three different CMV strains (AD169, Towne and TB40/E), whereas two antibodies against the AD-1 region of gB displayed minor neutralizing capabilities. In conclusion, the methods investigated in this proof-of-concept study enables for a semi-high-throughput workflow in the screening and investigation of biological active antibodies. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2225787
- author
- Carlsson, Fredrika LU ; Trilling, Mirko ; Perez, Franck and Ohlin, Mats LU
- organization
- publishing date
- 2012
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Immunological Methods
- volume
- 376
- issue
- Online 01 Dec 2011
- pages
- 69 - 78
- publisher
- Elsevier
- external identifiers
-
- wos:000301206800007
- scopus:84856556889
- pmid:22154743
- ISSN
- 1872-7905
- DOI
- 10.1016/j.jim.2011.11.010
- language
- English
- LU publication?
- yes
- id
- 5992623f-bb06-4aa3-bb52-d16c06bf14f6 (old id 2225787)
- date added to LUP
- 2016-04-01 14:13:55
- date last changed
- 2022-01-27 23:34:16
@article{5992623f-bb06-4aa3-bb52-d16c06bf14f6, abstract = {{Abstract in Undetermined<br/>Cytomegalovirus (CMV) causes severe sequelae in congenitally infected newborns and may cause life-threatening disease in immuno-deficient patients. Recent findings demonstrate the possibility to alleviate the disease by infusing intravenous immunoglobulin G (IgG) preparations, indicating that antibodies are an effective therapeutic option. Modern molecular methodologies, like phage display, allow for the development of specific antibodies targeting virtually any antigen, including those of CMV. However, such methodologies do not in general result in products that by themselves mediate biological activity. To facilitate a semi-high-throughput approach for functional screening in future efforts to develop efficacious antibodies against CMV, we have integrated two different approaches to circumvent potential bottlenecks in such efforts. Firstly, we explored an approach that permits easy transfer of antibody fragment encoding genes from commonly used phage display vectors into vectors for the production of divalent immunoglobulins. Secondly, we demonstrate that such proteins can be applied in a novel reporter-based neutralization assay to establish a proof-of-concept workflow for the generation of neutralizing antibodies against CMV. We validated our approach by showing that divalent antibodies raised against the antigenic domain (AD)-2 region of gB effectively neutralized three different CMV strains (AD169, Towne and TB40/E), whereas two antibodies against the AD-1 region of gB displayed minor neutralizing capabilities. In conclusion, the methods investigated in this proof-of-concept study enables for a semi-high-throughput workflow in the screening and investigation of biological active antibodies.}}, author = {{Carlsson, Fredrika and Trilling, Mirko and Perez, Franck and Ohlin, Mats}}, issn = {{1872-7905}}, language = {{eng}}, number = {{Online 01 Dec 2011}}, pages = {{69--78}}, publisher = {{Elsevier}}, series = {{Journal of Immunological Methods}}, title = {{A dimerized single-chain variable fragment system for the assessment of neutralizing activity of phage display-selected antibody fragments specific for cytomegalovirus.}}, url = {{http://dx.doi.org/10.1016/j.jim.2011.11.010}}, doi = {{10.1016/j.jim.2011.11.010}}, volume = {{376}}, year = {{2012}}, }