diff --git a/AutomatedTesting/Gem/PythonTests/Prefab/CMakeLists.txt b/AutomatedTesting/Gem/PythonTests/Prefab/CMakeLists.txt
index 48c24d1ebf..629db72dc7 100644
--- a/AutomatedTesting/Gem/PythonTests/Prefab/CMakeLists.txt
+++ b/AutomatedTesting/Gem/PythonTests/Prefab/CMakeLists.txt
@@ -12,7 +12,7 @@ if(PAL_TRAIT_BUILD_TESTS_SUPPORTED AND PAL_TRAIT_BUILD_HOST_TOOLS)
         NAME AutomatedTesting::PrefabTests
         TEST_SUITE main
         TEST_SERIAL
-        PATH ${CMAKE_CURRENT_LIST_DIR}/TestSuite_Main.py
+        PATH ${CMAKE_CURRENT_LIST_DIR}/TestSuite_Main_Optimized.py
         RUNTIME_DEPENDENCIES
             Legacy::Editor
             AZ::AssetProcessor
diff --git a/AutomatedTesting/Gem/PythonTests/Prefab/TestSuite_Main_Optimized.py b/AutomatedTesting/Gem/PythonTests/Prefab/TestSuite_Main_Optimized.py
new file mode 100644
index 0000000000..bf67b2c661
--- /dev/null
+++ b/AutomatedTesting/Gem/PythonTests/Prefab/TestSuite_Main_Optimized.py
@@ -0,0 +1,52 @@
+"""
+Copyright (c) Contributors to the Open 3D Engine Project.
+For complete copyright and license terms please see the LICENSE at the root of this distribution.
+
+SPDX-License-Identifier: Apache-2.0 OR MIT
+"""
+
+import pytest
+
+from ly_test_tools.o3de.editor_test import EditorSingleTest, EditorSharedTest, EditorParallelTest, EditorTestSuite
+
+
+@pytest.mark.SUITE_main
+@pytest.mark.parametrize("launcher_platform", ['windows_editor'])
+@pytest.mark.parametrize("project", ["AutomatedTesting"])
+class TestAutomationNoAutoTestMode(EditorTestSuite):
+
+    # Enable only -BatchMode for these tests. Some tests cannot run in -autotest_mode due to UI interactions
+    global_extra_cmdline_args = ["-BatchMode"]
+
+    class test_CreatePrefab_UnderAnEntity(EditorSharedTest):
+        from .tests.create_prefab import CreatePrefab_UnderAnEntity as test_module
+
+    class test_CreatePrefab_UnderAnotherPrefab(EditorSharedTest):
+        from .tests.create_prefab import CreatePrefab_UnderAnotherPrefab as test_module
+
+    class test_DeleteEntity_UnderAnotherPrefab(EditorSharedTest):
+        from .tests.delete_entity import DeleteEntity_UnderAnotherPrefab as test_module
+
+    class test_DeleteEntity_UnderLevelPrefab(EditorSharedTest):
+        from .tests.delete_entity import DeleteEntity_UnderLevelPrefab as test_module
+
+    class test_ReparentPrefab_UnderAnotherPrefab(EditorSharedTest):
+        from .tests.reparent_prefab import ReparentPrefab_UnderAnotherPrefab as test_module
+
+    class test_DetachPrefab_UnderAnotherPrefab(EditorSharedTest):
+        from .tests.detach_prefab import DetachPrefab_UnderAnotherPrefab as test_module
+
+    class test_OpenLevel_ContainingTwoEntities(EditorSharedTest):
+        from .tests.open_level import OpenLevel_ContainingTwoEntities as test_module
+
+    class test_CreatePrefab_WithSingleEntity(EditorSharedTest):
+        from .tests.create_prefab import CreatePrefab_WithSingleEntity as test_module
+
+    class test_InstantiatePrefab_ContainingASingleEntity(EditorSharedTest):
+        from .tests.instantiate_prefab import InstantiatePrefab_ContainingASingleEntity as test_module
+
+    class test_DeletePrefab_ContainingASingleEntity(EditorSharedTest):
+        from .tests.delete_prefab import DeletePrefab_ContainingASingleEntity as test_module
+
+    class test_DuplicatePrefab_ContainingASingleEntity(EditorSharedTest):
+        from .tests.duplicate_prefab import DuplicatePrefab_ContainingASingleEntity as test_module
\ No newline at end of file
diff --git a/AutomatedTesting/Gem/PythonTests/Prefab/tests/detach_prefab/DetachPrefab_UnderAnotherPrefab.py b/AutomatedTesting/Gem/PythonTests/Prefab/tests/detach_prefab/DetachPrefab_UnderAnotherPrefab.py
index 5c97f0b032..69dbcfc89c 100644
--- a/AutomatedTesting/Gem/PythonTests/Prefab/tests/detach_prefab/DetachPrefab_UnderAnotherPrefab.py
+++ b/AutomatedTesting/Gem/PythonTests/Prefab/tests/detach_prefab/DetachPrefab_UnderAnotherPrefab.py
@@ -7,8 +7,8 @@ SPDX-License-Identifier: Apache-2.0 OR MIT
 
 def DetachPrefab_UnderAnotherPrefab():
 
-    CAR_PREFAB_FILE_NAME = 'car_prefab'
-    WHEEL_PREFAB_FILE_NAME = 'wheel_prefab'
+    CAR_PREFAB_FILE_NAME = 'car_prefab2'
+    WHEEL_PREFAB_FILE_NAME = 'wheel_prefab2'
 
     import editor_python_test_tools.pyside_utils as pyside_utils