Exclusion criteria 1. Patients who have a history of hypersensitivity to the testing drug ingredients. 2. Patients with a previous history of gene therapy or any history of antitumour therapy in the last 4 weeks. 3. Female patients who are currently pregnant, suspect- ed of being pregnant, or expect to become pregnant during the study period. 4. Patients who are judged to be inappropriate as study subjects. 5. Patients who lack sufficient tumour tissues and periph- eral blood samples for gene sequencing. Termination criteria Patients can request withdrawal from the clinical trial at any time. If it becomes difficult to administrate the testing drug to the subject due to the following reasons, the responsible physician or the sharing physician will decide to discontinue the drug administration. 1. Any time it is found that the patient does not meet the inclusion criteria or meets the exclusion criteria. 2. Uncontrollable T cell proliferation. 3. SAEs, regardless of whether they are related to treat- ment. 4. Tumour progression. Study intervention and timeline of the study Individualised tumour peptide mixtures of 20 antigenic peptides will be injected subcutaneously for therapeutic Figure 1 Design flow chart of neoantigens. HLA, human leucocyte antigen; QC, quality control. 4 Wang L, et al. BMJ Open 2022;12:e055742. doi:10.1136/bmjopen-2021-055742Open access intervention in patients. The benchmark dose is 0.3 mg for each antigen peptide, and the single injection dose is 0.3 mg ×20=6.0 mg. Granulocyte- macrophage colony stimulating factor (GM- CSF) will be selected as a polypep- tide adjuvant, with the dosage of 0.5 μg/kg/dose. The medicine combine individualised tumour antigen polypeptides and GM- CSF will be injected subcutaneously at the lower edge of the upper arm deltoid muscle. The immunotherapy injection cycle is divided into two phases: initial immunisation injection and booster immunisation injection, which are completed in days 1, 4, 8, 15, 22 and weeks 12, 20. During the initial injection of individualised neoantigen tumour peptides, AEs will be recorded on the day of injection. During booster immunisation, AEs will be recorded for three consecutive days after each injec- tion. The efficacy of immunotherapy will be evaluated at 4, 8, 12, 24, 36 and 48 weeks. The timeline is displayed in figure 2. The individual follow- up for each participant is 12 months. Patients may enter in a succeeding phase (extended follow- up) according to the preset standards. The trial started in Q3/2019 and will last about 5 years. Assessment of therapeutic safety and efficacy After treatment with individual neoantigen tumour poly- peptides, all drug- related AEs occurring within 24 weeks will be observed and recorded. Adverse reactions that require close attention during the trial include cytokine storm- related acute inflammatory response syndrome, allergic reactions, neurotoxicity and systemic infections. PFS, ORR and DOR will be evaluated by CT/MRI according to the NCI criteria.30 Quality of life will be assessed using the Quality- of- Life Questionnaire of the European Organisation for Research and Treatment of Cancer 30.31 In addition, 2 mL peripheral blood samples will be collected and IFN-γ, CD4+, CD8 +antigen signals in peripheral blood will be detected by flow cytometry to evaluate the effectiveness of tumour neoantigen poly- peptide in inducing specific anti- tumour immunity. The changes of IFN-γ cells, CD4 +T cells, and CD8 +T cells in the peripheral blood mononuclear cell before and after immunotherapy will be analysed with ELISPOT technology. The efficacy evaluation also includes ctDNA detection before and after neoantigen immunotherapy. The observed indicators also include physical exam- ination, vital signs, Eastern Cooperative Oncology Group score, weight, blood routine, urine routine, liver and kidney function, coagulation function and tumour markers. At weeks 8, 12, 24, 36 and 48 of the study periods, patients’ target lesions will be scanned using enhanced CT or MRI to evaluate the therapeutic effect. The inter - vention and evaluation schedule are shown in table 1. Sample size To the best of our knowledge, there are no objective data on the efficacy of individualised neoantigens in advanced tumours. This study will not stipulate a sample size a priori. The sample size is not therefore determined by power calculations. The purpose of the trial is to evaluate the safety of individualised neoantigens, similar to the phase I clinical trials of drugs, so our sample size is deter - mined to be 20 patients. Statistical considerations First, we need to list the number of selected and completed cases, confirm three data sets: full analysis set, per- protocol set, safety data set and then describe demo- graphic data and other baseline characteristic values. Descriptive data analysis will be used to evaluate the experimental data. The percentage of subjects with medi- cation compliance greater than 80%, and the percentage of subjects with combination medication will be calcu- lated. AEs will be coded according to International Conference on Harmonisation (ICH) Dictionary of International Medical Terms (MedDRA). List AE, SAE, major AE, AE leading to disengagement and calculate the incidence. Data from patients who received at least five sessions of individualised tumour neoantigen polypeptide Figure 2 The study flow chart. GM- CSF , granulocyte- macrophage colony- stimulating factor. 5 Wang L, et al.