University System of Maryland

The United States Government As Represented by the Department of Veterans Affairs

Mohammad Sajadi

George K Lewis

Anthony DeVico

Yongjun Guan

Marzena Pazgier

Tongyun Liu

Dongkyoon Kim

Guy Cavet

The present invention provides broadly neutralizing antibodies against HIV, compositions comprising the same and methods of use thereof.

Broadly neutralizing antibodies against HIV

The present invention provides a method of making an antibody by identifying a circulating antibody with activity from a subject comprising i) subjecting biological fluid selected from the group consisting of blood, plasma and serum and combinations thereof from the subject to one or more rounds of affinity chromatography to purify the circulating antibody; ii) optionally further subjecting the circulating antibody to isoelectric focusing to purify the circulating antibody based on charge; iii) testing the purified circulating antibody for activity; iv) digesting the purified circulating antibody from parts i) or ii) to create an antibody fragment; v) subjecting the antibody fragment to mass spectrometry to generate a mass assignment and a deduced amino acid sequence of the antibody fragment; vi) comparing the deduced amino acid sequence with an amino acid sequence of an antibody generated from the subject's B-cells to identify an antibody sequence that matches the deduced amino acid sequence; vii) generating an antibody comprising light chain and heavy chain CDR sequences of the B-cell antibody that matches the deducted amino acid sequence of party vi); and viii) testing the antibody of part vii) for activity.

Methods of making active antibodies from biological fluids

Fusion proteins comprising a portion of the HIV-1 Env protein (gp120 and gp140) and single-chain fragment V regions (ScFv) of an enhancing antibody that exhibits binding specificity for HIV-1 Env protein are disclosed that may serve in immunogenic formulations for vaccination against HIV-1 infection, as well as methods of generating an immune response using the fusion proteins.

Recombinant fusion protein comprising HIV gp120 linked to an enhancing CD4 binding site mAb

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Cockeysville, MD, United States of America

Mohammad Sajadi