Article: Use of recombinant human antibody fragments for detection of cytomegalovirus antigenemia

The determination and quantitation of peripheral blood leukocytes (PBLs) expressing human cytomegalovirus (HCMV) antigens is widely employed in clinical virology for rapid diagnosis of HCMV-related infections. We describe how CMV antigenemia may be accurately detected by means of human recombinant monoclonal Fab fragments rescued from a combinatorial phage display library prepared from an HCMV-infected donor. Fourteen recombinant Fabs were tested against HCMV-positive PBLs from a patient with ongoing HCMV infection. Three clones were found to react specifically with the nuclei of these cells. These three recombinant Fabs were subsequently tested, individually and pooled together, against 60 PBL samples taken from immunosuppressed patients. The reactivity observed was comparable to that obtained with mouse monoclonal antibodies commercially available for this purpose. The three recombinant Fabs were shown to react specifically with the 65-kDa viral tegument phosphoprotein encoded by UL83 (pUL83), which is the most abundant viral antigen in HCMV-infected PBLs.

Citing Articles

Understanding differences between synthetic and natural antibodies can help improve antibody engineering.

Burkovitz A, Ofran Y MAbs. 2015; 8(2):278-87.

PMID: 26652053 PMC: 4966605. DOI: 10.1080/19420862.2015.1123365.

Production and characterization of a human recombinant monoclonal Fab fragment specific for influenza A viruses.

Desogus A, Burioni R, Ingianni A, Bugli F, Pompei R, Fadda G Clin Diagn Lab Immunol. 2003; 10(4):680-5.

PMID: 12853404 PMC: 164267. DOI: 10.1128/cdli.10.4.680-685.2003.

Characterization of a type-common human recombinant monoclonal antibody to herpes simplex virus with high therapeutic potential.

De Logu A, Williamson R, Rozenshteyn R, Ramiro-Ibanez F, Simpson C, Burton D J Clin Microbiol. 1998; 36(11):3198-204.

PMID: 9774565 PMC: 105301. DOI: 10.1128/JCM.36.11.3198-3204.1998.

References

1.

Rand K, Houck H, Dickinson M . False positive indirect fluorescent antibody (IFA) tests for antibody to herpes simplex virus 1. Comparison of four commercially available methods. Am J Clin Pathol. 1986; 86(6):765-8. DOI: 10.1093/ajcp/86.6.765. View

2.

van der Bij W, Torensma R, van Son W, Anema J, Schirm J, Tegzess A . Rapid immunodiagnosis of active cytomegalovirus infection by monoclonal antibody staining of blood leucocytes. J Med Virol. 1988; 25(2):179-88. DOI: 10.1002/jmv.1890250208. View

3.

Revello M, Zavattoni M, Percivalle E, Grossi P, Gerna G . Correlation between immunofluorescent detection of human cytomegalovirus immediate early antigens in polymorphonuclear leukocytes and viremia. J Infect Dis. 1989; 160(1):159-60. DOI: 10.1093/infdis/160.1.159. View

4.

van den Berg A, Klompmaker I, Haagsma E, Bijleveld C, Schirm J, VAN DER GIESSEN M . Antigenemia in the diagnosis and monitoring of active cytomegalovirus infection after liver transplantation. J Infect Dis. 1991; 164(2):265-70. DOI: 10.1093/infdis/164.2.265. View

5.

Barbas 3rd C, Kang A, Lerner R, Benkovic S . Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc Natl Acad Sci U S A. 1991; 88(18):7978-82. PMC: 52428. DOI: 10.1073/pnas.88.18.7978. View

6.

Kain S, Adams M, Kondepudi A, Yang T, Ward W, Kitts P . Green fluorescent protein as a reporter of gene expression and protein localization. Biotechniques. 1995; 19(4):650-5. View

7.

Landini M . New approaches and perspectives in cytomegalovirus diagnosis. Prog Med Virol. 1993; 40:157-77. View

8.

Williamson R, Burioni R, Sanna P, Partridge L, Barbas 3rd C, Burton D . Human monoclonal antibodies against a plethora of viral pathogens from single combinatorial libraries. Proc Natl Acad Sci U S A. 1993; 90(9):4141-5. PMC: 46462. DOI: 10.1073/pnas.90.9.4141. View

9.

Burioni R, Williamson R, Sanna P, Bloom F, Burton D . Recombinant human Fab to glycoprotein D neutralizes infectivity and prevents cell-to-cell transmission of herpes simplex viruses 1 and 2 in vitro. Proc Natl Acad Sci U S A. 1994; 91(1):355-9. PMC: 42946. DOI: 10.1073/pnas.91.1.355. View

10.

Burton D, Barbas 3rd C . Human antibodies from combinatorial libraries. Adv Immunol. 1994; 57:191-280. DOI: 10.1016/s0065-2776(08)60674-4. View

11.

Gerna G, Revello M, Percivalle E, Morini F . Comparison of different immunostaining techniques and monoclonal antibodies to the lower matrix phosphoprotein (pp65) for optimal quantitation of human cytomegalovirus antigenemia. J Clin Microbiol. 1992; 30(5):1232-7. PMC: 265256. DOI: 10.1128/jcm.30.5.1232-1237.1992. View

Use of Recombinant Human Antibody Fragments for Detection of Cytomegalovirus Antigenemia