Engineered Antibody Expression Constructs

The isolated RT22 VH gene was subcloned in-frame, without additional amino acids, at Not I/Apa I sites of pcDNA 3.4–RT11-HC carrying the human IgG1 constant-domain sequence (CH1-hinge-CH2-CH3) with a mutation of CH3 at three sites (L234A, L235A, and P329G) for HC expression. For LC expression, the CT03 VL gene was subcloned in-frame, without additional amino acids, at Not I/Bsi WI sites of pcDNA 3.4–TMab4-LC carrying the human κ constant domain sequence (residues 108 to 214). For LC expression of integrin αvβ5/αvβ3–targeting cyclic peptide–fused antibodies, the synthesized DNA encoding the cyclic peptides (RGD10 and in4) and a six–amino acid linker (MGSSSN) was subcloned in-frame, without additional amino acids, at the N terminus of CT03 VL via the Not I/Bsi WI sites of pcDNA 3.4–CT03-LC. All the constructs were confirmed by sequencing (Macrogen, Korea).

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Shin S.M., Kim J.S., Park S.W., Jun S.Y., Kweon H.J., Choi D.K., Lee D., Cho Y.B, & Kim Y.S. (2020). Direct targeting of oncogenic RAS mutants with a tumor-specific cytosol-penetrating antibody inhibits RAS mutant–driven tumor growth. Science Advances, 6(3), eaay2174.

Publication 2020

Amino acids Antibodies Cyclic peptides Frame Gene Human Igg1 Integrin αvβ3 Mutation

Corresponding Organization : Ajou University

Other organizations : Samsung Medical Center, Sungkyunkwan University

Top 5 similar protocols

Protocol cited in 4 other protocols

Variable analysis

independent variables

Expression of the isolated RT22 VH gene
Expression of the CT03 VL gene
Expression of the synthesized DNA encoding the cyclic peptides (RGD10 and in4) and a six–amino acid linker (MGSSSN)

dependent variables

Not explicitly mentioned

control variables

Subcloning of the RT22 VH gene in-frame, without additional amino acids, at Not I/Apa I sites of pcDNA 3.4–RT11-HC carrying the human IgG1 constant-domain sequence (CH1-hinge-CH2-CH3) with a mutation of CH3 at three sites (L234A, L235A, and P329G) for HC expression
Subcloning of the CT03 VL gene in-frame, without additional amino acids, at Not I/Bsi WI sites of pcDNA 3.4–TMab4-LC carrying the human κ constant domain sequence (residues 108 to 214)
Subcloning of the synthesized DNA encoding the cyclic peptides (RGD10 and in4) and a six–amino acid linker (MGSSSN) in-frame, without additional amino acids, at the N terminus of CT03 VL via the Not I/Bsi WI sites of pcDNA 3.4–CT03-LC

positive controls

Not mentioned

negative controls

Not mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!