Therapeutic antibodies for treatment and prophylaxis of transmittable viral diseases
a technology for transmittable diseases and therapeutic antibodies, applied in the field of treatment and prevention of transmittable diseases, can solve the problems of lack of protection, limited or no options, and inability to detect antibodies, and achieve the effect of high neutralization titer
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example 1
Preparation of High Neutralization Titer Serum for Viral Protective Bird Antibody
[0058] Antibody titer measured by serum neutralization (SN) assays were performed to provide analyses of the protective capabilities of the goose antibody to viral infection over traditional ELISA assays that measure binding affinity to viral epitopes. Such traditional methods have shown discrepancies in the past. For example, in studies performed by Ben-Nathan et al. in obtaining antibodies from human subjects, the ELISA titer was reported to be 1:1600; however, the functional protective capabilities were shown to be substantially lower and were actually in the range of 1:80 to 1:320 (J Infect Dis 2003; 188: 5-12).
[0059] In the present study, geese of a variety of ages were exposed to West Nile Virus, sera were collected from the geese, and the sera of the infected geese were tested using a sera microtiter neutralization plaque assay to measure the usefulness of the sera for protecting cells from vir...
example 2
Evaluation of Goose Antiserum for Presence of West Nile Virus
[0062] Goose antiserum was examined for the presence of West Nile Virus RNA by RT-PCR analysis. DNA was amplified from the prepared RNA in a Perkin-Elmer Model 480 thermal cycler. Primers were designed to map the conserved sequences of the polyprotein gene (West Nile Virus Oligo Detect Kit, WNV Primer Mix [Part No. 5653], Chemicon International, California). The RT-PCR was performed with the QIAGEN one-step RT-PCR kit (QIAGEN, Valencia, Calif.) by using 5 μl of RNA and 0.3 μM of each primer in a 50 μl total reaction volume following the manufacture's protocol. When the PCR mixture was complete, the samples were overlaid with two drops of molecular biology grade mineral oil. All previous manipulations were performed in a Nuaire biological safety cabinet Model NU 425-400. The following cycling times and temperatures were used: cDNA synthesis; 50° C. for 30 minutes, 94° C. for 15 minutes followed by 40 cycles of 94° C. for 1...
example 3
Purification of Goose Antibodies to West Nile Virus
[0065] Twenty liters of sera collected from geese infected with West Nile Virus was irradiated for 67 minutes / 300 ml aliquots to eliminate any residual virus present in the sera, and the samples were examined by polymerase chain reaction (PCR) to ensure that the sera was virus free. The antibody fraction of the sera was purified by density centrifugation, dialyzed to remove gradient, and concentrated to approximately 3 times the original protein concentration. Purity of the goose antibody was established using RT-PCR analysis. All preparations were greater than 1:4000 determined by a microtiter plaque neutralization assay.
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