Evaluation of different lymphoid tissue sources for the construction of human immunoglobulin gene libraries.
BACKGROUND
Phage display technology allows the isolation of novel human monoclonal antibodies. The technology relies on the construction of a recombinant antibody library and its display on phage particles. The quality of an antibody library is affected by several factors including the size, diversity and source of immunoglobulin genes.


OBJECTIVE
The aim of the project was to determine the best tissue source for the construction of antibody libraries.


METHODS
Three tissue sources were used in this study: peripheral blood mononuclear cells from a healthy donor, Epstein-Barr virus (EBV) transformed peripheral blood mononuclear cells and lymph node tissue from individuals with breast cancer. The quality of each tissue source was assessed using two criteria: (1) the number of mature and activated B cells in each source; (2) the amount of immunoglobulin heavy and light chain genes amplifiable by polymerase chain reaction (PCR).


RESULTS
EBV-transformed peripheral blood mononuclear cells and lymph node tissue were shown to contain more B cells than peripheral blood mononuclear cells. A relatively larger amount of immunoglobulin gene products could be amplified from EBV-transformed peripheral blood mononuclear cells and the lymph node. However, immunoglobulin containing gamma 1 chains could not be amplified from EBV-transformed mononuclear cells, and the resultant pattern of gene amplification suggests a possible selection bias.


CONCLUSIONS
This study indicates that among the three tissue sources examined, lymph node tissue is the most suitable source for the construction of antibody libraries.
Yip
Y L
YL
Department of Biotechnology, University of NSW, Sydney, Australia.
Hawkins
N J
NJ
Clark
M A
MA
Ward
R L
RL
D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics; D002472:Cell Transformation, Viral; D005434:Flow Cytometry; D015723:Gene Library; D005803:Genes, Immunoglobulin; D004854:Herpesvirus 4, Human; D006801:Humans; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D008214:Lymphocytes / Q000378:metabolism / Q000502:physiology; D008221:Lymphoid Tissue / Q000378:metabolism / Q000502:physiology*; D010641:Phenotype; D016133:Polymerase Chain Reaction
D000911:Antibodies, Monoclonal