Cytotoxic immunoglobulin
a technology of cytotoxic immunoglobulin and immunoglobulin, which is applied in the field of cytotoxic immunoglobulin, can solve the problems of limitation of use of such monovalent phages, single fusion proteins, and monovalent phage displays, and achieve the effect of small size and easy penetration through cell layers or tumors
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
Problems solved by technology
Method used
Image
Examples
example 1
Construction of the Non-Focussed Fcab Library (Fcab0D and Phage Surface Display
[0256]The crystal structure of an IgGI Fc fragment, which is published in the Brookhaven Database as entry 1 OQO.pdb was used to aid in the design of the Fcab library.
[0257]The sequence which was used as the basis for construction of the Fcab library is given in SEQ ID No.1 (FIG. 3). In this sequence, the first amino acid corresponds to GIu 216 of human IgGI (EU numbering; according to the IMGT database (imgt.cines.fr / textes / IMGTrepertoire / Proteins / protein / human / IGH / IGHC / Hu_IGHC allgenes.html; lookup 2007 Jun. 25), it is the first residue of the human IgGI hinge region, which is given as: (E)PKSCDKTHTCPPCP) (SEQ ID NO:441) of the heavy constant chain hinge region of human IgGI.) The second-last residue of SEQ ID No.1 (FIG. 3) corresponds to GIy 446 of human IgGI (EU numbering; IMGT: residue number 129 of the CH3 domain of human IgG1).
[0258]After detailed analysis of the structure of loqo.pdb and by visual...
example 2
Construction of the Focused Fcab Library (Fcab02) and Phage Surface Display
[0263]As described in example 1, an Fcab library was prepared in which the randomized library positions are fully randomized, i.e. they are encoded by a codon such as NNS, NNB, NNK, NNN or others are used.
[0264]For clarity, the meaning of the letters such as N, B, S or K is defined by the IUPAC nucleotide ambiguity code, which is given in the following table:
TABLE 1IUPAC nucleotide ambiguity codeSymbolMeaningNucleic AcidAAAdenineCCCytosineGGGuanineTTThymineUUUracilMA or CRA or GWA or TSC or GYC or TKG or TVA or C or GHA or C or TDA or G or TBC or G or TXG or A or T or CNG or A or T or C
[0265]Source: Nomenclature for incompletely specified bases in nucleic acid sequences: recommendations 1984. A Cornish-Bowden, Nucleic Acids Res. 1985 May 10; 13(9): 3021-3030.
[0266]These codons given above are designed such that all 20 amino acids are encoded by them. It may be preferable to choose subsets out of the possible ...
example 3
Construction of a Phage Surface Display Library with Additional Amino Acid Residues Between the Library Insert (Binding Partner) and p3
[0269]In order to investigate accessibility of the potential binding site of the displayed protein a binding assay is performed: the phage suspension is reacted with anti-myc mAb 9E10-coated microplates (or immunotubes). After washing, the bound phages are detected with anti-M13-enzyme conjugate. As a control, helper phage—which does not display the protein fusion and the myc-tag is reacted with the plates. Other controls are reaction of phages with non-coated plates and reaction of phages with antiserum recognizing the p3-fusion partner of the phages.
[0270]Ideally, the anti-myc-reactivity of phages displaying the p3-fusion protein should give very clear ELISA readouts whereas helper phage reactions to anti-myc-mAb should not be above background (non-coated plates). The structure of a CH3 dinner displayed at the surface of an M13 phage through bindin...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More