Anti-IL13 human antibodies
An antibody, L-CDR3 technology, applied in the field of those molecules that neutralize IL-13 activity, which can solve problems such as lack of signaling motifs, short cytoplasmic tails, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0149] Preparation of Transfectomas Producing Monoclonal Antibodies
[0150] Antibodies of the invention can also be produced in host cell transfectomas using, for example, a combination of recombinant DNA techniques and gene transfection methods well known in the art (eg, Morrison, S. (1985) Science 229:1202).
[0151] For example, to express an antibody, or an antibody fragment thereof, DNA encoding partial or full-length light and heavy chains can be obtained by standard molecular biology techniques (e.g., PCR amplification or cDNA cloning using hybridomas expressing the antibody of interest) , and the DNA can be inserted into an expression vector such that the gene is operably linked to transcriptional and translational control sequences. In this context, the term "operably linked" means that the antibody gene is ligated into a vector such that the transcriptional and translational control sequences within the vector perform their intended function of regulating the tran...
Embodiment 1
[0209] Example 1: Generation of human IL-13-specific antibodies from an immunized spleen library
[0210] RNA from the spleen was used to generate a phage display library of H and L chain variable domains randomly distributed in a Fab phage display vector as described in US Patent 6,794,132. Phage display libraries were subjected to five rounds of selection using biotinylated hrIL-13 in a solution phase equilibrium binding protocol as described in the patent. The first four rounds choose to use 10 -8 M's hrIL-13, last round of selection using 10 -9 hrIL-13 of M. The final signal-to-noise ratio for this library, determined by calculating the pfu recovered in the presence of antigen divided by the pfu recovered in its absence, was 37, indicating that greater than 90% of the selected phage express antibodies that bind hrIL-13. This phage display library was then subcloned into a plasmid vector for expression of soluble Fab as described in US Patent 6,794,132. The subcloned li...
Embodiment 2
[0211] Example 2: Quantitative analysis of binding affinity: identification of anti-human IL-13 Fab candidates
[0212] Surface plasmon resonance measurements were performed using an optical biosensor BIAcore 2000, which quantifies the interaction of anti-IL-13 Fab with some hrIL-13. The specific binding of IL-13 to the respective IL-13 Fab immobilized on the BIAcore chip can be measured by following the accumulation of the ligand on the receptor. Microscopic binding (k on ) and dissociation rate (k off ) can be obtained directly from the on-chip mass accumulation rate and expressed in response units (RU). Anti-IL-13 Fab was immobilized on the chip surface by secondary anti-human LK antibody (Jackson Immunochemicals). The capture antibody was covalently bound using the 'Amine coupling kit' (BIAcore, Cat. No. BR-1000-50) as recommended by the manufacturer's protocol. 250 μl of different concentrations of hrIL-13 were injected at a flow rate of 20 μl / min and kinetic traces w...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More