TopSpec platform (pI-trap/Orbitrap/Omnitrap) combined data

Data 1 name: 20221221_500nulInflix250ngulNIST_180umcell-90minfocusing_2_externalpower_alwayson_omnitrap_ECD40ms_2 Description: A 4 µL mixture of infliximab (500ng/µL) and NISTmAb (250ng/µL) is aspirated by the Alias autosampler pI-trap system and transferred into the pI cell for 90-min, 2000V cIEF separation. After the focusing, the sample is slowly injected at 0.5 µL/min to Exploris 480 – omnitrap. Two experiments are set in the data acquisition method: 1. MS full scan with resolution 120 000. 2&3. MS/MS experiment performed by orbi-omnitrap. Two m/z ranges, 2481 ± 5 representing infliximab and 2471 ± 5 representing NISTmAb, respectively, are isolated and injected to the omnitrap, where ECD is executed with filament current 5.65A and irradiation time 40ms. Infliximab is the earlier peak at RT 12 -14 min, NISTmAb is the later peak 22 - 24 min. Data 2 name: 20221221_500nulInflix250ngulNIST_180umcell-90minfocusing_2_externalpower_alwayson_omnitrap_HCD30_1 Description: A 4 µL mixture of infliximab (500ng/µL) and NISTmAb (250ng/µL) is aspirated by the Alias autosampler pI-trap system and transferred into the pI cell for 90-min, 2000V cIEF separation. After the focusing, the sample is slowly injected at 0.5 µL/min to Exploris 480 – omnitrap. Three experiments are set in the data acquisition method: 1. MS full scan with resolution 120 000. 2&3. MS/MS experiment performed by orbi-omnitrap. Two m/z ranges, 2481 ± 5 representing infliximab and 2471 ± 5 representing NISTmAb, respectively, are isolated and get fragmented with HCD by orbitrap. HCD collision energy is set at 30%. Infliximab is the earlier peak at RT 12 -14 min, NISTmAb is the later peak 22 - 24 min. Data 3 name: 20221216_1ugulInflix100ngulNIST_180umcell-90minfocusing_2_externalpower_alwayson_omnitrap_EID50ms_2 Description: A 4 µL mixture of infliximab (1000ng/µL) and NISTmAb (100ng/µL) is aspirated by the Alias autosampler pI-trap system and transferred into the pI cell for 90-min, 2000V cIEF separation. After the focusing, the sample is slowly injected at 0.5 µL/min to Exploris 480 – omnitrap. Two experiments are set in the data acquisition method: 1. MS full scan with resolution 60 000. 2&3. MS/MS experiment performed by orbi-omnitrap. Two m/z ranges, 2481 ± 5 representing infliximab and 2471 ± 5 representing NISTmAb, respectively, are isolated and injected to the omnitrap, where ECD is executed with filament current 5.65A and irradiation time 40ms. 4. MS full scan with resolution 7500. Infliximab is the earlier peak at RT 12 -14 min, NISTmAb is the later peak 22 - 24 min.数据集1 名称：20221221_500nulInflix250ngulNIST_180umcell-90minfocusing_2_externalpower_alwayson_omnitrap_ECD40ms_2 描述：取4微升由英夫利昔单抗（infliximab，500ng/µL）与NIST单抗（NISTmAb，250ng/µL）组成的混合样品，经Alias自动进样器（autosampler）等电点捕获（pI-trap）系统吸取后，转移至等电点池（pI cell）中，于2000V电压下进行90分钟的毛细管等电聚焦（capillary isoelectric focusing, cIEF）分离。聚焦完成后，以0.5 µL/min的流速将样品缓慢注入Exploris 480-离子阱（omnitrap）质谱系统。数据采集方法设置了两组实验：1. 分辨率为120000的质谱全扫描（MS full scan）；2、3. 由轨道离子阱（orbi-omnitrap）完成的串联质谱（MS/MS）实验。分别选取对应英夫利昔单抗的质荷比（mass-to-charge ratio, m/z）窗口2481 ± 5，以及对应NIST单抗的质荷比窗口2471 ± 5进行离子分离，并注入离子阱中，以5.65A的灯丝电流、40ms的辐照时长执行电子捕获解离（electron capture dissociation, ECD）操作。英夫利昔单抗的色谱峰出峰更早，保留时间（retention time, RT）为12~14分钟；NIST单抗的色谱峰出峰较晚，保留时间为22~24分钟。 数据集2 名称：20221221_500nulInflix250ngulNIST_180umcell-90minfocusing_2_externalpower_alwayson_omnitrap_HCD30_1 描述：取4微升由英夫利昔单抗（infliximab，500ng/µL）与NIST单抗（NISTmAb，250ng/µL）组成的混合样品，经Alias自动进样器等电点捕获系统吸取后，转移至等电点池中，于2000V电压下进行90分钟的毛细管等电聚焦分离。聚焦完成后，以0.5 µL/min的流速将样品缓慢注入Exploris 480-离子阱质谱系统。数据采集方法设置了三组实验：1. 分辨率为120000的质谱全扫描；2、3. 由轨道离子阱完成的串联质谱实验。分别选取对应英夫利昔单抗的质荷比窗口2481 ± 5，以及对应NIST单抗的质荷比窗口2471 ± 5进行离子分离，随后通过轨道阱以高能碰撞解离（high-energy collisional dissociation, HCD）模式进行碎裂，碰撞能量设置为30%。英夫利昔单抗的色谱峰出峰更早，保留时间为12~14分钟；NIST单抗的色谱峰出峰较晚，保留时间为22~24分钟。 数据集3 名称：20221216_1ugulInflix100ngulNIST_180umcell-90minfocusing_2_externalpower_alwayson_omnitrap_EID50ms_2 描述：取4微升由英夫利昔单抗（infliximab，1000ng/µL）与NIST单抗（NISTmAb，100ng/µL）组成的混合样品，经Alias自动进样器等电点捕获系统吸取后，转移至等电点池中，于2000V电压下进行90分钟的毛细管等电聚焦分离。聚焦完成后，以0.5 µL/min的流速将样品缓慢注入Exploris 480-离子阱质谱系统。数据采集方法设置了两组实验：1. 分辨率为60000的质谱全扫描；2、3. 由轨道离子阱完成的串联质谱实验。分别选取对应英夫利昔单抗的质荷比窗口2481 ± 5，以及对应NIST单抗的质荷比窗口2471 ± 5进行离子分离，并注入离子阱中，以5.65A的灯丝电流、40ms的辐照时长执行电子捕获解离操作。4. 分辨率为7500的质谱全扫描。英夫利昔单抗的色谱峰出峰更早，保留时间为12~14分钟；NIST单抗的色谱峰出峰较晚，保留时间为22~24分钟。