Detection and typing of herpes simplex viruses by using recombinant immunoglobulin fragments produced in bacteria.
Thirty-seven bacterial clones producing human recombinant monoclonal antibody Fab fragments (rFabs) reactive to herpes simplex virus (HSV) antigens were selected from a human combinatorial antibody library constructed in a phage-display vector by a panning procedure against an HSV lysate. Thirty-four of the HSV-specific rFabs were able to specifically recognize HSV-infected cells in indirect immunofluorescence (IF) assays; of these, 25 recognized cells infected by either HSV type 1 (HSV-1) or HSV-2, while 9 recognized only HSV-1-infected cells. One HSV type-common rFab (rFab H37) and one HSV-1-specific rFab (rFab H85) were further evaluated as reagents for viral detection and typing by IF staining in 134 HSV-positive (72 HSV-1 and 62 HSV-2) viral cultures from clinical specimens. The results obtained with these two rFabs were fully consistent with those obtained with a commercial preparation of fluorescein-labeled anti-HSV type-specific murine monoclonal antibodies. The detection sensitivity with the type-common rFab in indirect IF assays was higher overall than that provided by the type-specific murine monoclonal antibodies. Preparations of rFabs suitable for IF staining can be easily and inexpensively obtained in a clinical microbiology laboratory from Escherichia coli cultures. Similar HSV-specific rFabs, therefore, could be advantageous for in vitro diagnostic purposes.
Cattani
P
P
Istituto di Microbiologia, Università Cattolica del Sacro Cuore, Rome,Italy.
Rossolini
G M
GM
Cresti
S
S
Santangelo
R
R
Burton
D R
DR
Williamson
R A
RA
Sanna
P P
PP
Fadda
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000235:genetics; D000956:Antigens, Viral / Q000032:analysis; D001483:Base Sequence; D002522:Chlorocebus aethiops; D004926:Escherichia coli / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000145:classification / Q000276:immunology; D018258:Herpesvirus 2, Human / Q000145:classification / Q000276:immunology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D017422:Sequence Analysis, DNA; D018139:Simplexvirus / Q000145:classification* / Q000276:immunology / Q000302:isolation & purification; D014709:Vero Cells
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011993:Recombinant Fusion Proteins