Diagnostics using conditionally active antibodies
A method of detecting tumor cells in a sample containing cells. In the method, a sample is encapsulated to produce a capsule with the sample encapsulated therein. The capsule containing the sample encapsulated therein is incubated under cell culture conditions suitable for supporting cell activity and growth of tumor cells and the encapsulated sample from the incubated capsule is contacted with a conditionally active antibody that has a higher binding affinity to a cell surface protein of a tumor cell under a first value of a condition of a tumor microenvironment, in comparison with the binding affinity of the conditionally active antibody to the same cell surface protein under a second value of the condition.
1. A method of detecting a presence of tumor cells having a cell surface protein in a sample containing cells, said method comprising steps of:
encapsulating the sample to produce one or more capsules with the sample encapsulated therein wherein the one or more capsules comprise a protein and a nutrient suitable for supporting cell activity and growth of the tumor cells;
incubating the one or more capsules with the sample encapsulated therein under cell culture conditions suitable for supporting cell activity and growth of the tumor cells whereby the one or more incubated capsules containing said tumor cells produce a first value of a condition of a tumor microenvironment selected from the group consisting of a pH in a range of 6.0-6.8, a partial oxygen pressure in a range of from about 1 to about 20 mmHg and a glucose concentration in a range of from about 0.05 to about 0.5 mM in said capsules containing said tumor cells;
contacting the encapsulated sample in the one or more incubated capsules with a conditionally active antibody that has a detectably higher binding affinity to the cell surface protein under the first value of the condition of the tumor microenvironment selected from the group consisting of a pH in a range of 6.0-6.8, a partial oxygen pressure in a range of from about 1 to about 20 mmHg and a glucose concentration in a range of from about 0.05 to about 0.5 mM, in comparison with a binding affinity of the conditionally active antibody to the same cell surface protein under a second value of the condition selected from the group consisting of a pH in a range of 7.0-7.8, a partial oxygen pressure in a range of from about 30 to about 50 mmHg, and a glucose concentration in a range of from about 2.5 to about 10 mM; and
detecting the presence of tumor cells by detecting the detectably higher binding affinity of the conditionally active antibody to the cell surface protein at the first value of the condition of the tumor microenvironment selected from the group consisting of a pH in a range of 6.0-6.8, a partial oxygen pressure in a range of from about 1 to about 20 mmHg and a glucose concentration in a range of from about 0.05 to about 0.5 mM relative to binding affinity of the conditionally active antibody to the cell surface protein under the second value of the condition selected from the group consisting of a pH in a range of 7.0-7.8, a partial oxygen pressure in a range of from about 30 to about 50 mmHg, And a glucose concentration in a range of from about 2.5 to about 10 mM.
2. The method of claim 1 , wherein the one or more capsules comprise:
a core comprising the sample with the cells suspended therein; and
a shell comprising a biocompatible polymer.
3. The method of claim 2 , wherein the core comprises a polymer matrix.
4. The method of claim 2 , wherein the biocompatible polymer is selected from the group consisting of proteins and polysaccharides.
5. The method of claim 4 , wherein the proteins are selected from the group consisting of albumin, collagen, synthetic polyamino acids and prolamins.
6. The method of claim 4 , wherein the polysaccharides are selected from the group consisting of alginate, cellulose and heparin.
7. The method of claim 2 , wherein the biocompatible polymer is selected from the group consisting of poly(lactide), poly(glycolide), poly(lactide-co-glycolide), poly(caprolactone), polycarbonates, polyamides, polyanhydrides, polyphosphazene, polyamino acids, polyortho esters, polyacetals, polycyanoacrylates, biodegradable polyurethanes, polyacrylates, ethylene-vinyl acetate polymers, acyl-substituted cellulose acetates, polyurethanes, polystyrenes, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonated polyolefins, and polyethylene oxide.
8. The method of claim 2 , wherein the core comprises the protein and the nutrient suitable for supporting cell activity and growth of the tumor cells.
9. The method of claim 1 , wherein the one or more capsules have a mean diameter of from about 10 μm to about 1000 μm.
10. The method of claim 1 , wherein the incubating step is carried out for a period of at least about 3 hours to about 36 hours.
11. The method of claim 1 , wherein the conditionally active antibody is conjugated to a detectable label.
12. The method of claim 11 , wherein the detectable label is a molecule or an ion directly or indirectly detectable based on light absorbance, fluorescence, reflectance, light scatter, phosphorescence, luminescence properties, radioactive properties, nuclear magnetic resonance properties or paramagnetic properties.
13. The method of claim 1 , wherein the cell surface protein is a protein product of a housekeeping gene.
14. The method of claim 13 , wherein the housekeeping gene is selected from the group consisting of AP2S1, CD81, GPAA1, LGALS9, MGAT2, MGAT4B, VAMP3, SLC2A1, SLC2A2, SLC2A3, SLC2A4, SLC2A5, SLC2A6, SLC2A7, SLC2A8, SLC2A9, SLC2A10, SLC2A11, SLC2A12, SLC2A13, and SLC2A14.
15. The method of claim 1 , wherein the cell surface protein is selected from the group consisting of ABCA7, ABCC1, ABCC5, ABHD3, ACKR3, ADAM10, AQP1, AQP3, ATP13A3, ATP1B3, ATP2B1, ATP2B4, ATP6AP1, ATP6V0A2, BACE1, BMPR2, BNIP2, BST2, BTN2A1, BTN3A3, C12orf76, C17orf62, C1orf27, CCDC107, CD4, CD44, CD46, CD81, CD9, CD99L2, CDAN1, CDIPT, CLCN6, CNNM4, CYP20A1, DCBLD2, DHRS7B, ERBB2, ETNK1, FAM210B, GINM1, GPI, GRAMD1A, HELZ, HERPUD1, HMOX1, HPS3, ICAM1, IFI30, IFRD1, IL15RA, IL6ST, ITGA7, ITGB1, ITGB4, ITGB5, ITSN1, JAG1, LAIR1, LMTK2, LRBA, LRP12, LSR, MACF1, MADD, MCAM, MCOLN1, MET, MICAL3, MPV17L2, NCKIPSD, NDC1, NEO1, NOTCH2, PANX1, PDLIM5, PFDN1, PGAP3, PGRMC2, PHLDB2, PIGN, PIGQ, PIGW, PKN2, PTPRS, PVR, RALGAPA2, RNF145, RNF149, SC5D, SCAMP4, SDC2, SDC4, SLC12A2, SLC16A1, SLC16A3, SLC17A5, SLC19A1, SLC1A5, SLC30A1, SLC38A6, SLC38A7, SLC39A14, SLC3A2, SLC43A1, SLC46A1, SLC46A3, SLC4A2, SLC4A7, SLC7A5, SLC9A1, SMAGP, SORT1, SPG11, SPINT2, SPPL2B, SPPL3, SRD5A3, SRPRB, STX18, STX4, SYVN1, TAPT1, TAZ, TBC1D5, TGFBR2, TM2D2, TMEM183A, TMEM205, TMEM218, TMEM222, TMEM245, TMEM258, TMEM50A, TMEM63B, TMEM97, TNFRSF12A, TXNDC11, UBR2, UQCC1, VSIG4, WWP1, YIPF4, ZDHHC20 and ZDHHC5.
16. The method of claim 1 , further comprising a step of expanding a population of the cells in the sample.
17. The method of claim 1 , further comprising a step of enriching a population of the cells in the sample.
18. The method of claim 1 , wherein the binding affinity of said conditionally active antibody at the first value of the condition in the tumor microenvironment is at least 5 fold higher than the binding affinity of the conditionally active antibody at the second value of the condition.