Method For Generating and Selecting Antibodies Against Target Protein
Compositions are provided that comprise antibody against membrane proteins such as chemokine receptors. In particular, monoclonal human antibodies against human CXCR4 are provided that are capable of inhibiting HIV infection and chemotaxis in human breast cancer cells. The antibodies can be used as prophylactics or therapeutics to prevent and treat HIV infection and cancer, for screening drugs, and for diagnosing diseases or conditions associated with interactions with chemokine receptors.
1 . An antibody that binds to loop 6 of human CXCR4.
2 . The antibody of claim 1 , wherein the antibody is a monoclonal antibody.
3 . The antibody of claim 2 , wherein the monoclonal antibody is a single chain antibody.
4 . The antibody of claim 3 , wherein the single chain antibody is encoded by DNA SEQ ID NO: 59 or 62.
5 . The antibody of claim 2 , wherein the heavy chain variable region of the monoclonal antibody comprises an amino acid sequence with SEQ ID NO: 60 or 63.
6 . The antibody of claim 2 , wherein the light chain variable region of the monoclonal antibody comprises an amino acid sequence with SEQ ID NO: 61 or 64.
7 . The antibody of claim 1 , wherein the loop 6 of human CXCR4 comprises SEQ ID NO: 58.
8 . The antibody of claim 1 , wherein the antibody is capable of inhibiting infection of human immunodeficiency virus (HIV).
9 . The antibody of claim 8 , wherein the HIV is HIV type I or HIV type 2.
10 . The antibody of claim 1 , wherein the antibody is capable of inhibiting chemotaxis of cancer cells.
11 . The antibody of claim 10 , wherein the cancer cells are selected from the group consisting of breast, prostate, liver, renal, lung, skin, ovarian, cervical, brain, thyroid, stomach, colon, lymphoma, leukemia and pancreas cancer cells.
12 . A recombinant expression vector encoding a polypeptide selected from the group consisting of SEQ ID NOs: 60, 61, 63, and 64.
13 . The recombinant expression vector of claim 12 , wherein the vector is a bacterial, yeast, plant, mammalian or viral expression vector.
14 . A recombinant cell expressing a polypeptide selected from the group consisting of SEQ ID NOs: 60, 61, 63, and 64.
15 . The recombinant cell of claim 14 , wherein the cell is a bacterial, yeast, plant or mammalian cell.
16 . The recombinant cell of claim 15 , wherein the cell is a human cell.
17 . A method for preventing or treating HIV infection, comprising:
administering the antibody of claim 1 to a human.
18 . The method of claim 17 , further comprising:
administering to the host an anti-retroviral agent.
19 . The method of claim 18 , wherein the anti-retroviral agent is selected from the group consisting of nucleoside and non-nucleoside HIV reverse transcriptase inhibitors, HIV protease inhibitors, and HIV integrase inhibitors.
20 . The method of claim 19 , wherein the nucleoside HIV reverse transcriptase inhibitor is selected from the group consisting of zidovudine, didanosine, zalcitabine, lamivudine, stavudine, abacavir, and adefovir dipivoxil.
21 . The method of claim 19 , wherein the non-nucleoside HIV reverse transcriptase inhibitor is selected from the group consisting of nevirapine, delavirdine and efavirenz.
22 . The method of claim 19 , wherein the HIV protease inhibitor is selected from the group consisting of indinavir, ritonavir, saqinavir, nelfinavir, and amprenavir.
23 . A method for preventing or treating cancer, comprising:
administering the antibody of claim 1 to a human.
24 . The method of claim 23 , further comprising:
administering an anti-neoplastic agent to the human.
25 . The method of claim 24 , wherein the anti-neoplastic agent is selected from alkylating agents, antibiotic agents, antimetabolic agents, hormonal agents, plant-derived agents, and biologic agents.
26 . The method of claim 25 , wherein the alkylating agent is cyclophosphamide or cisplatin.
27 . The method of claim 25 , wherein the antibiotic agent is doxorubicin, mitomycin C, or bleomycin.
28 . The method of claim 25 , wherein the antimetabolic agent is fluorouracil or methotrexate.
29 . The method of claim 25 , wherein the hormonal agent is tamoxifen.
30 . The method of claim 25 , wherein the plant-derived agent is paclitaxel, vincristine or camptothecin.
31 . The method of claim 25 , wherein the biologic agent is Trastruzumab, Rituximab, or a cytokine.