Antibodies and chimeric antigen receptors specific for CD19
Provided are CD19 binding molecules, including anti-CD19 antibodies, including antibody fragments such as single-chain fragments, and chimeric receptors including the antibodies, such as chimeric antigen receptors (CARs). Among the antibodies are human antibodies, including those that compete for binding to CD19 with reference antibodies, such as murine antibodies. In some embodiments, the antibodies display similar functional properties to the reference antibodies, such as comparable binding affinities and/or competitive inhibition properties. Also provided are genetically engineered cells expressing the chimeric receptors, and uses of the binding molecules and cells adoptive cell therapy.
1 . A conjugate, comprising:
a) an anti-CD19 antibody or antigen-binding fragment thereof, comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein:
the VH region comprises a heavy chain complementarity determining region 1 (CDR-H1), a CDR-H2, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NOs: 18, 19, and 20, respectively, and
the VL region comprises a light chain complementarity determining region 1 (CDR-L1), a CDR-L2, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NOs: 21, 22, and 23, respectively; SEQ ID NOs: 21, 22, and 24, respectively; SEQ ID NOs: 25, 26, and 27, respectively; SEQ ID NOs: 28, 29, and 30, respectively; SEQ ID NOs: 31, 32, and 33, respectively; SEQ ID NOs: 74, 94, and 103, respectively; SEQ ID NOs: 73, 93, and 101, respectively; SEQ ID NOs: 75, 95, and 104, respectively; SEQ ID NOs: 76, 96, and 105, respectively; or SEQ ID NOs: 73, 93, and 102, respectively; and
b) a cytotoxic agent, an imaging agent, an enzyme or a fragment thereof, a detectable moiety, or a multimerization domain.
2 . The conjugate of claim 1 , wherein the conjugate comprises the anti-CD19 antibody or antigen-binding fragment thereof and a cytotoxic agent.
3 . The conjugate of claim 2 , wherein the cytotoxic agent is selected from the group consisting of chemotherapeutic agents or drugs, growth inhibitory agents, enzymes, toxins, and radioactive isotopes.
4 . The conjugate of claim 2 , wherein the cytotoxic agent is a chemotherapeutic agent or drug selected from the group consisting of a maytansinoid, an auristatin, a dolastatin, a calicheamicin or derivative thereof, an anthracycline, methotrexate, vindesine, a taxane, a trichothecene, CC1065, and a vinca alkyloid.
5 . The conjugate of claim 4 , wherein:
(a) the auristatin is monomethylauristatin drug moieties DE or DF (MMAE or MMAF); and/or
(b) the anthracycline is daunomycin or doxorubicin; and/or
(c) the taxane is selected from the group consisting of docetaxel, paclitaxel, larotaxel, tesetaxel, and ortataxel; and/or
(d) the vinca alkyloid is selected from the group consisting of vincristine, vinblastine, and etoposide.
6 . The conjugate of claim 2 , wherein the cytotoxic agent is a chemotherapeutic agent or drug selected from the group consisting of methotrexane, adriamicin, vincristine, vinblastine, etoposide, doxorubicin, melphalan, mitomycin C, chlorambucil, and daunorubicin.
7 . The conjugate of claim 2 , wherein the cytotoxic agent is a toxin or fragment thereof selected from the group consisting of diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain, ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins, Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, neomycin, and tricothecenes.
8 . The conjugate of claim 2 , wherein the cytotoxic agent is a radioactive isotope selected from the group consisting of At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 , and radioactive isotopes of Lu.
9 . The conjugate of claim 1 , wherein the conjugate comprises the anti-CD19 antibody or antigen-binding fragment thereof and an imaging agent.
10 . The conjugate of claim 9 , wherein the imaging agent is a radionuclide.
11 . The conjugate of claim 10 , wherein the radionuclide is selected from the group consisting of 111 In, 99 Tc, 14 C, 131 I, 125 I, and 3 H.
12 . The conjugate of claim 1 , wherein the conjugate comprises the anti-CD19 antibody or antigen-binding fragment thereof and an enzyme or a fragment thereof.
13 . The conjugate of claim 12 , wherein the enzyme or a fragment thereof is a nucleolytic enzyme.
14 . The conjugate of claim 1 , wherein the conjugate comprises the anti-CD19 antibody or antigen-binding fragment thereof and a detectable moiety.
15 . The conjugate of claim 14 , wherein the detectable moiety is selected from the group consisting of radionuclides, enzymes, fluorescent moieties or proteins, and luminescent moieties.
16 . The conjugate of claim 14 , wherein the detectable moiety is a radionuclide that is a radioactive isotope of chromium, cobalt, fluorine, gadolinium, germanium, holmium, indium, iodine, lanthanium, lutetium, manganese, molybdenum, palladium, phosphorous, praseodymium, promethium, rhenium, rhodium, rutheroium, samarium, scandium, selenium, strontium, sulfur, technetium, thallium, tin, tritium, xenon, ytterbium, and yttrium.
17 . The conjugate of claim 14 , wherein the detectable moiety is a radionuclide selected from the group consisting of 51 Cr, 57 Co, 18 F, 153 Gd, 159 Gd, 68 Ge, 166 Ho, 115 In, 113 In, 112 In, 111 In, 125 I, 131 I, 123 I, 121 I, 140 La, 177 Lu, 54 Mn, 99 Mo, 103 Pd, 32 P, 142 Pr, 149 Pm, 186 Re, 188 Re, 105 Rh, 97 Ru, 153 Sm, 47 Sc, 75 Se, 85 Sr, 35 S, 99 Tc, 201 Ti, 113 Sn, 117 Sn, 3 H, 133 Xe, 169 Yb, 175 Yb, and 90 Y.
18 . The conjugate of claim 14 , wherein the detectable moiety is an enzyme selected from the group consisting of alkaline phosphatase, horseradish peroxidase, luciferase, and β-galactosidase.
19 . The conjugate of claim 14 , wherein the detectable moiety is a fluorescent moiety or protein selected from the group consisting of fluorescein, rhodamine, phycoerythrin, green fluorescent protein (GFP), and blue fluorescent protein (BFP).
20 . The conjugate of claim 14 , wherein the detectable moiety is a luminescent moiety that is a nanoparticle.