Universal method for detecting total concentration of monoclonal antibody drug of targeted soluble protein in monkey serum
A monoclonal antibody, general-purpose technology, applied in the direction of color/spectral characteristic measurement, analysis by causing chemical reaction of materials, material analysis by observing the influence on chemical indicators, etc., can solve time-consuming cost and reagent cost , method development failure, method performance not up to standard and other problems, to achieve the effect of reducing time cost and reagent cost, ensuring accuracy and reliability, and significant economic benefits
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Embodiment 1
[0034] A general-purpose enzyme-linked immunoassay method for detecting the total concentration of IgG4 monoclonal antibody drugs targeting complement C5 protein in cynomolgus monkey serum:
[0035] 1. Main test steps
[0036] Coating: the mouse anti-human IgG monoclonal antibody that can specifically bind to human IgG1, IgG2, IgG3 and IgG4 subtypes was prepared with pH9.4-pH9.8 carbonate buffer to prepare 2μg / mL coating antibody Add 100 μL of working solution to each well, seal the plate with a sealing film, and incubate at 2-8°C for 14-20 hours;
[0037] Plate washing: wash the microplate plate 3 times with 0.05% PBST (1×PBS buffer containing 0.05% Tween 20), 300 μL / well, without soaking, after the last wash, pat the microplate plate on absorbent paper Dry;
[0038] Sealing: Add 250 μL of 0.05% PBST containing 0.2% I-Block to each well, seal the plate with a sealing film, place at 37±1°C, and incubate at 400 rpm for 1h±5min;
[0039] Plate washing: wash the plate with 0.0...
Embodiment 2
[0064] A general-purpose enzyme-linked immunoassay method for detecting the total concentration of IgG2 monoclonal antibody drugs targeting complement C5 protein in cynomolgus monkey serum:
[0065] 1. Main test steps
[0066] The main test steps are the same as in Example 1 except that the prepared concentrations of the standard curve calibration standard sample and the quality control substance are different.
[0067] 2. Experimental results
[0068] 1) Standard curve result
[0069] In this example, the standard curve contains calibration standards at 9 concentration levels: 200000.000ng / mL, 160000.000ng / mL, 80000.000ng / mL, 40000.000ng / mL, 20000.000ng / mL, 10000.000ng / mL, 5000.000ng / mL mL, 2500.000ng / mL, 1250.000ng / mL; 200000.000ng / mL is the upper limit of quantification (ULOQ), 2500.000ng / mL is the lower limit of quantitation (LLOQ), and 1250.000ng / mL is the anchor point. The calibration standard sample was prepared from the mixed serum of cynomolgus monkeys used for med...
Embodiment 3
[0083] A general-purpose enzyme-linked immunoassay method for detecting the concentration of completely free monoclonal antibody drugs targeting an IgG1 type monoclonal antibody drug targeting an unknown membrane protein in cynomolgus monkey serum:
[0084] 1. Main test steps
[0085] The main test steps are the same as those in Example 1 except that the number of standard samples calibrated by the standard curve, the prepared concentration and the prepared concentration of the quality control substance are different.
[0086] 2. Experimental results
[0087] 1) Standard curve result
[0088] In this example, the standard curve contains calibration standards at 8 concentration levels: 50000.000ng / mL, 40000.000ng / mL, 20000.000ng / mL, 10000.000ng / mL, 6000.000ng / mL, 3000.000ng / mL, 1500.000ng / mL mL, 750.000ng / mL; 50000.000ng / mL is the upper limit of quantification (ULOQ), 1500.000ng / mL is the lower limit of quantitation (LLOQ), and 750.000ng / mL is the anchor point. The calibrati...
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