Antibody Generation and Purification

Antibodies from QA255 were originally isolated and cloned as described previously [65 (link)]. In brief, paired heavy and light chain DNA clones were co-transfected in equal ratios into 293F cells (293 Freestyle cells; Thermo Fisher; 1x10⁶ cells/1 μg of total DNA) with a 16:1:1 (OptiPRO Serum-Free Medium:293Max:DNA, Thermo Fisher) ratio. Antibodies were harvested after 72 h and purified using Protein G resin in hand-packed, gravity flow columns (Pierce). Antibody concentration was determined using protein absorbance at 280 nM (Nanodrop).

Free full text: Click here

Williams K.L., Stumpf M., Naiman N.E., Ding S., Garrett M., Gobillot T., Vézina D., Dusenbury K., Ramadoss N.S., Basom R., Kim P.S., Finzi A, & Overbaugh J. (2019). Identification of HIV gp41-specific antibodies that mediate killing of infected cells. PLoS Pathogens, 15(2), e1007572.

Publication 2019

293 Freestyle cells

Antibodies Antibody Cells Clones Gravity Light Protein Protein g Resin Serum

Corresponding Organization : Fred Hutch Cancer Center

Other organizations : University of Washington, Université de Montréal, Stanford University

Top 5 similar protocols

Variable analysis

independent variables

Ratios of heavy and light chain DNA clones co-transfected into 293F cells

dependent variables

Antibody concentration
Antibody harvesting after 72 h

control variables

Cell line (293F cells)
Transfection reagents (OptiPRO Serum-Free Medium, 293Max)
Total DNA amount (1 μg) per 1x10^6 cells

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly 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!