Human antibodies, pharmaceutical compositions and methods
A technology of antibodies and carbohydrates, applied in drug combinations, chemical instruments and methods, antibodies, etc., can solve problems such as limitations of applicability and binding characteristics
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Embodiment 1
[0284] Embodiment 1: clinical sample collection
[0285] To generate human monoclonal antibodies against Globo series antigens, B cells were isolated from the peripheral blood of vaccinated patients. After administration of Globo H-KLH vaccine (OBI-822 / OBI-821 ) in recurrent ovarian cancer patients, blood samples were collected for the following analytical procedures.
Embodiment 2
[0286] Example 2: Sorting and culturing of human single B cells
[0287] IgD freshly isolated from human peripheral blood - IgM - IgA - Memory B cells and dispensed into 384-well tissue culture plates at a density of 1 cell per well using a fluorescence-activated cell sorter. Sorted B cells were stimulated to secrete IgG and incubated for several days. After incubation, B cell lysates and culture supernatants were collected separately.
Embodiment 3
[0288] Example 3: Amplification of antibody genes
[0289] In obtaining Globo H, SSEA-3 or SSEA-4 binding clones, genes encoding Ig VH, IgVκ or IgVλ were recovered from B cell lysates using RT-PCR and cloned into IgG expression vectors. Recombinant antibodies are expressed by transfection of mammalian cells and used to confirm binding specificity or perform other functional assays.
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