الملخص

تعد الأضداد وحيدة النسيلة (mAb) العارية والمرتبطة مع دواء (ADC) فئة مهمة جدًا من الأدوية البيولوجية. يعد تقييم مرتسم تغاير الشحنات احدى الأدوات المستخدمة لضمان جودة هذه الفئة العلاجية. كان الهدف الرئيسي من الدراسة الحالية هو تقييم تأثير تغيير بعض شروط الكروماتوغرافيا البؤرية المعتمدة على استخدام أعمدة مبادلة للأيونات السالبة على مرتسم الشحنات لضد وحيد النسيلة عارٍ أو مرتبط مع دواء. شملت الشروط الكروماتوغرافية التي تم التحقق منها: حجوم ما قبل الشطف المدروج، درجة حرارة العمود، pH المحاليل الدارئة البادئة والمستخدمة للشطف، وإضافة محاليل أمفوليت حاملة ذات المجال 8-10.5. أظهرت نتائج الكروماتوغرافيا البؤرية أن mAb-1 مكون من نظيرين لهما قيمpI  مساوية 8.1 و8.2. عند ربط mAb-1 الى دواء سام للخلايا أظهرت النتائج الكروماتوغرافية  قمم اكثر للضد المرتبط موافقة لقيم pI من 7.3 إلى 8.5. تم اختبار انتقائية الطريقة عن طريق تحليل ضد آخر mAb-2. أظهر mAb-2 تجانساً وقلوية اكبر من mAb-1  مع قمة رئيسية موافقة لقيم .8.7 pI طبقت الشروط الكروماتوغرافية البؤرية الأمثل لمراقبة ثبات الضد المرتبط المدروس الخاضع لشروط ثبات قاسية.  اثبتت الطريقة الكروماتوغرافية البؤرية المطورة امكانية استخدامها لإظهار تغيرات في مرتسم مغايرات الشحنة سواء لضد عارٍ او مرتبط مع دواء.

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كلمات مفتاحية : الأجسام المضادة، الأضداد المرتبطة، نظائر مختلفة الشحنات، نقطة التعادل الكهربائي، الكروماتوغرافيا البؤرية.

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