Korean J Pediatr Infect Dis.  1999 Nov;6(2):253-260. 10.14776/kjpid.1999.6.2.253.

Development of Monoclonal Antibody to Cytomegalovirus Major Immediate-early(α) Protein

• Park JH
• Sohn YM
• Park KH
• Lee WY

• KMID: 2440264
• DOI: http://doi.org/10.14776/kjpid.1999.6.2.253

Abstract

PURPOSE
The purpose of this study is to make and use monoclonal Ab which reacts with CMV major immediate early(α) protein(p72).
METHODS
Normal human fibroblast(Foreskin derived) was cultured in Eagle's minimal essential medium(MEM) containing 10% cowfetus serum and mouse chondroblast was cultured in P3X63 Ag8.653(ATCC, Maryland USA) to maintain 5×10#x2075;/m1 cell counts. CMV(KJHJ90) from congenital CMV infected infant's urine was multiplied and used for Ab making. CMV Ag was injected 4 times, 1 week interval into the peritoneal space of 6~8 weeks old mice. And then lymphocytes and fibroblasts taken from spleen were obtained and conjugated. After the conjugated cell cultured, we chose the cell that had high Ab titer using indirect immunofluerescent method.
RESULTS
Among the 28 monoclonal antibodies obtained LPC12 and LPC23 reacted highly with nucleus of AD169 infected cell. Purified AD169 after SDS-PAGE, molecular weight of Ag, which reacted with purified monoclonal Ab, was obtained using Western blotting. Monoclonal Ab of LPC12 and LPC23 clone reacted most highly with 72 kd Ag.
CONCLUSION
LPC12 and LPC23 clonal Ab with AD 169(P63-27) is useful on early diagnosis of CMV infection.

Keyword

Cytomegalovirus; Immediate early protein

MeSH Terms

Animals
Antibodies, Monoclonal
Blotting, Western
Cell Count
Chondrocytes
Clone Cells
Cytomegalovirus*
Early Diagnosis
Electrophoresis, Polyacrylamide Gel
Fibroblasts
Humans
Lymphocytes
Maryland
Methods
Mice
Molecular Weight
Spleen
Antibodies, Monoclonal